Inhibitors of dipeptidyl peptidase iv

ABSTRACT

Novel compounds that are inhibitors of one or most post-proline cleaving proteases, e.g. dipeptidyl peptidase IV, according to general formula (1). R 1  is H or CN, X 1  is O, S, CH 2 , CHF, CF 2 , CH(CH 3 ), C(CH 3 ) 2  or CH(CN), and b is 1 or 2. G 1  is H or a group according to the formula —CH 2 —X 2 —(CH 2 ) a —G 3  and G 2  is H or a group according to the formula —CH 2 —(CH 2 9 a —G 3 , provided that one of G 1  and G 2  is H and the other is not H. X 2  is O, S, or CH 2 , and a is 0, 1 or 2, provided that when a is 1 then X 2  is CH 2 . G 3  is a group according to one of general formulae 2-4, where the variables have meaning given in the description. The compounds are useful in the treatment of i.a. type 2 diabetes and impaired glucose tolerance.

The present invention relates to novel compounds that are inhibitors of post-proline aminopeptidases. The compounds are useful as antiproliferative agents and in the treatment of, inter alia, type 2 diabetes and impaired glucose tolerance.

BACKGROUND

The enzyme dipeptidyl peptidase IV, herein abbreviated DP-IV (and elsewhere as DAP-IV or DPP-IV) and also known by the classification EC.3.4.14.5, is a serine protease that cleaves the N-terminal dipeptide from peptides that begin with the sequence H-Xaa-Pro (where Xaa is any amino acid, although preferably a lipophilic one, and Pro is proline). It will also accept as substrates peptides that begin with the sequence H-Xaa-Ala (where Ala is alanine). DP-IV was first identified as a membrane-bound protein. More recently a soluble form has been identified.

Initial interest in DP-IV focussed on its role in the activation of T lymphocytes. DP-IV is identical to the T cell protein CD26. It was proposed that inhibitors of DP-IV would be capable of modulating T cell responsiveness, and so could be developed as novel immunomodulators. It was further suggested that CD26 was a necessary co-receptor for HIV, and thus that DP-IV inhibitors could be useful in the treatment of AIDS.

Attention was given to the role of DP-IV outside the immune system. It was recognised that DP-IV has a key role in the degradation of several peptide hormones, including growth hormone releasing hormone (GHRH) and glucagon-like peptide-1 and -2 (GLP-1 and GLP-2). Since GLP-1 is known to have a potentiating effect on the action of insulin in the control of post-prandial blood glucose levels it is clear that DP-IV inhibitors might also be usefully employed in the treatment of type II diabetes and impaired glucose tolerance. At least two DP-IV inhibitors are currently undergoing clinical trials to explore this possibility.

Several groups have disclosed inhibitors of DP-IV. While some leads have been found from random screening programs, the majority of the work in this field has been directed towards the investigation of substrate analogues. Inhibitors of DP-IV that are substrate analogues are disclosed in, for example, U.S. Pat. No. 5,462,928, U.S. Pat. No. 5,543,396, WO95/15309 (equivalent to U.S. Pat. No. 5,939,560 and EP 0731789), WO98/19998 (equivalent to U.S. Pat. No. 6,011,155), WO99/46272 and WO99/61431.

More recently a number of proteins have been found that share some of the enzymatic properties of DP-IV. Some, such as FAP and DPP-8, have sequence homology with DP-IV, while others, such as QPP, have no such homology but nevertheless mimic the aminodipeptidase activity of DP-IV. The physiological function of these newer proteases is still being investigated. FAP has been implicated in invasive processes such as cancer metastasis and endometriosis, and QPP appears to be involved in immune-cell apoptosis. It is also possible that some of these proteases share a common function. This redundancy would allow continuing normal physiological function in the event of a failure in the expression or function of one of the proteases.

In order to further define the roles of these newer proteases it is important to have the tools to manipulate selectively each one or the whole class. Therefore there exists a need for specific and potent inhibitors of each of these proteases, and also for potent non-specific inhibitors of the class of post-proline cleaving aminodipeptidases.

SUMMARY OF THE INVENTION

We disclose herein a series of novel compounds that are inhibitors of one or more post-proline cleaving proteases, and specifically compounds according to general formula 1.

In general formula 1, R¹ is H or CN, X¹ is O, S, CH₂, CHF, CF₂, CH(CH₃), C(CH₃)₂ or CH(CN), and b is 1 or 2. G¹ is H or a group according to the formula —CH₂—X²—(CH₂)_(a)—G³ and G² is H or a group according to the formula —CH₂—(CH₂)_(a)—G³, provided that one of G¹ and G² is H and the other is not H. X² is O, S or CH₂, and a is 0, 1 or 2, provided that when a is 1 then X² is CH₂. G³ is a group according to one of general formulae 2-4.

X³, X⁴ and X⁵ are either nitrogen N or CH, provided that at least two of X³, X⁴ and X⁵ are N. X⁶ is either O or NH. R² is either H or alkyl. R³ is selected from H, Cl, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂. R⁴, R⁵, R⁶, R⁷ and R⁸ are selected from H, Br, Cl, F, CF₃, alkyl, acyl, OH, O-alkyl, NH₂, NH-alkyl, N(alkyl)₂, NO₂, NH-acyl, CO₂H, CO₂-alkyl, CONH₂, CONH-alkyl, CON(alkyl)₂ and CN. X⁷ is CH₂, O, S or NH. R⁹ is either H or alkyl. R¹⁰, R¹¹, R¹², R¹³ and R¹⁴ are selected from H, Br, Cl, F, CF₃, alkyl, acyl, OH, O-alkyl, NH₂, NH-alkyl, N(alkyl)₂, NO₂, NH-acyl, CO₂H, CO₂-alkyl, CONH₂, CONH-alkyl, CON(alkyl)₂ and CN. R¹⁵ and R¹⁶ are each independently H, alkyl, alkenyl, polyfluoroalkyl, aralkyl, aryl or CH₂—L—R¹⁷, where L is a covalent bond, CH═CH, C≡C or —C₆H₄—, and R¹⁷ is H, alkyl or aryl, or R¹⁵ and R¹⁶ together are a group according to one of general formulae 5-7.

R¹⁸ is H, alkyl, aryl, OH, O-alkyl, NH₂, NH-alkyl or N(alkyl)₂, and R¹⁹ is H, alkyl, aryl, F, Cl, Br, CF₃, OH, O-alkyl, NH₂, NH-alkyl or N(alkyl)₂. The integers d and e are 0, 1, 2 or 3 such that d+e is 3, 4 or 5, and f is 1, 2 or 3. When R¹⁵ and R¹⁶ are both H then X¹ may not be S or CH₂ if b is 1.

Preferred compositions are inhibitors of non-membrane associated post-proline cleaving proteases. The most preferred compositions are selective for non-membrane associated proteases (e.g. for example inhibitors of one or more of QPP, DPP-8 and/or DPP-9).

DETAILED DESCRIPTION OF THE INVENTION

In a first aspect, the present invention relates to a series of novel α-amino acyl derivatives of saturated nitrogen-containing heterocycles according to general formula 1.

In general formula 1, the group R¹ is either a hydrogen atom H or a nitrile group CN. The group X¹ is selected from an oxygen atom O, a sulphur atom S, a methylene group CH₂, a monofluoromethylene group CHF, a difluoromethylene group CF₂, an ethylidene group CH(CH₃), a 2-propylidene group C(CH₃)₂ and a cyanomethylene group CH(CN). The integer b is either 1 or 2, such that the nitrogen-containing ring has 5 or 6 members.

The group G¹ is either H or a group according to the formula —CH₂—X²—(CH₂)_(a)—G³ and the group G² is either H or a group according to the formula —CH₂—(CH₂)_(a)—G³, provided that one of G¹ and G² is H and the other is not H. The group X² is selected from O, S and CH₂. The integer a is 0, 1 or 2, provided that when a is 1 then X² is CH₂.

The group G³ is selected from a group according to general formula 2, a group according to general formula 3 and a group according to general formula 4.

In general formula 2, the groups X³, X⁴ and X⁵ are selected from nitrogen N and methine CH, provided that at least two of X³, X⁴ and X⁵ are nitrogen. Preferably X³, X⁴ and X⁵ are all nitrogen. The group X⁶ is selected from O and NH. R² is selected from H and alkyl. R³ is selected from H, Cl, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂. R⁴, R⁵, R⁶, R⁷ and R⁸ are independently selected from H, Br, Cl, F, CF₃, alkyl, acyl, OH, O-alkyl, NH₂, NH-alkyl, N(alkyl)₂, NO₂, NH-acyl, CO₂H, CO₂-alkyl, CONH₂, CONH-alkyl, CON(alkyl)₂ and CN.

In general formula 3, the group X⁷ is selected from CH₂, O, S and NH. R⁹ is selected from H and alkyl. R¹⁰, R¹¹, R¹², R¹³ and R¹⁴ are independently selected from H, Br, Cl, F, CF₃, alkyl, acyl, OH, O-alkyl, NH₂, NH-alkyl, N(alkyl)₂, NO₂, NH-acyl, CO₂H, CO₂-alkyl, CONH₂, CONH-alkyl, CON(alkyl)₂ and CN.

In general formula 4, R¹⁵ and R¹⁶ are each independently selected from H, alkyl, alkenyl, polyfluoroalkyl, aralkyl, aryl and CH₂—L—R¹⁷, where L is selected from a covalent bond, CH═CH, C≡C and —C₆H₄— and R¹⁷ is selected from H, alkyl and aryl, or R¹⁵ and R¹⁶ together are a group selected from general formula 5, general formula 6 and general formula 7.

In these general formulae, the group R¹⁸ is selected from H, alkyl, aryl, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂, and the group R¹⁹ is selected from H, alkyl, aryl, F, Cl, Br, CF₃, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂. The integers d and e are selected from 0, 1, 2 and 3 such that d+e is 3, 4 or 5, and the integer f is selected from 1, 2 and 3.

When R¹⁵ and R¹⁶ are both H then X¹ may not be S or CH₂ if b is 1.

The term alkyl, as used herein, denotes saturated hydrocarbon groups with between 1 and 10 carbon atoms, including straight-chain, branched and mono- and polycycloalkyl groups, such as methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, cyclopentyl, cyclohexylmethyl, 2-cyclohexyl-2-propyl, bicyclo[2.2.2]octyl and the like.

The term alkenyl, as used herein, denotes monounsaturated hydrocarbon groups with between 2 and 10 carbon atoms, including straight-chain, branched and mono- and polycycloalkenyl groups, such as vinyl, allyl, methallyl, cyclohex-3-enyl and the like.

The term aryl, as used herein, denotes monocyclic and fused bicyclic aromatic groups, including carbocyclic groups, such as phenyl and naphthyl, and heteroaryl groups with up to three heteroatoms selected from nitrogen, oxygen and sulphur, such as pyrrolyl, furyl, thienyl, pyrazolyl, imidazolyl, oxazolyl, isothiazolyl, pyridyl, pyrimidinyl, indolyl, quinolinyl and the like. Unless otherwise specified, aryl groups may optionally be substituted with up to three groups independently selected from alkyl, OH, O-alkyl, Cl, F, Br, NH₂, NH-alkyl, N(alkyl)₂, CO₂H, CO₂-alkyl, CONH₂, CONH-alkyl, CON(alkyl)₂, NO₂ and CN.

The term aralkyl, as used herein, denotes alkyl groups that are substituted by, or fused to, one or more aryl groups, including benzyl, phenethyl, indanyl, fluorenyl and the like.

The term acyl, as used herein, denotes a group selected from H—CO, alkyl-CO, aryl-CO and aralkyl-CO, including formyl, acetyl, benzoyl, phenylacetyl and the like.

The term polyfluoroalkyl, as used herein, denotes an alkyl group wherein all the hydrogen atoms on one or more of the carbon atoms are replaced by fluorine atoms, including trifluoromethyl, 2,2,2-trifluoroethyl and the like.

In one preferred embodiment of the invention R¹ is H.

In another preferred embodiment of the invention R¹ is CN.

In another preferred embodiment of the invention X¹ is CH₂.

In another preferred embodiment of the invention X¹ is S.

In another preferred embodiment of the invention b is 1.

In another preferred embodiment of the invention b is 2.

In another preferred embodiment of the invention a is 0.

In another preferred embodiment of the invention a is 0 and X² is CH₂.

In another preferred embodiment of the invention a is 1.

In another preferred embodiment of the invention a is 1 and X² is CH₂.

In another preferred embodiment of the invention a is 2 and X² is CH₂.

In another preferred embodiment of the invention the compound is a compound according to general formula 8.

In another preferred embodiment of the invention the compound is a compound according to general formula 9.

In another preferred embodiment of the invention the compound is a compound according to general formula 10.

In another preferred embodiment of the invention the compound is a compound according to general formula 11.

In another preferred embodiment of the invention the compound is a compound according to general formula 12.

In another preferred embodiment of the, invention the compound is a compound according to general formula 13.

It will be recognised that certain of the compounds within the scope of the present invention are capable of forming salts with suitable acids or bases. To the extent that such salts are pharmaceutically acceptable they are included within the scope of this invention

It will further be recognised that certain of the compounds within the scope of the present invention are capable of existing as optical isomers, such as enantiomers and diastereomers. All such optical isomers and mixtures thereof, including but not limited to racemates, are included within the scope of the invention.

The compounds of the present invention are inhibitors of post-proline cleaving proteases such as DPP-IV, QPP, FAP, DPP-8 (DPRP-1) and DPP-9 (DPRP-2). As such they may be useful in the treatment of diseases in which dysregulation of these enzymes or their endogenous substrates plays a role or the disease is ameliorated by inhibition of such enzymes. Accordingly, in further aspects, the present invention provides for the use of compounds according to the present invention in the preparation of pharmaceutical compositions, and for the use of such compositions a therapeutic agents.

Preferred compositions which are inhibitors for QPP may have G²=H, b=1 or 2 and/or a=0 or 1. Further preferred compositions having b=2 include G1 groups having a=0 or 1 and X² is CH₂. Further preferred compositions having b=2 have X¹=CH₂ or S, for example Example 38 of Table 2. Further preferred compositions having b=1 include G1 groups having a=0 or 1 and X² is CH₂. Further preferred compositions having b=1 have X¹=S or CH₂ or CF₂, for example, Example 42 of Table 2.

The compounds of the present invention can be prepared by methods generally known in the art and illustrated in the following non-limiting examples.

EXAMPLES Example 1 (2S)-1-[N^(ω),N^(ω)-(Dicinnamyl)-L-lysinyl]pyrrolidine-2-carbonitrile dihydrochloride

A. (N^(α)-(tert-Butyloxycarbonyl)-N^(ω)-(9-fluorenylmethyloxycarbonyl)-L-lysinyl)-L-prolinamide

N^(ω)-(tert-Butyloxycarbonyl)-N^(ω)-(9-fluorenylmethyloxycarbonyl)-L-lysine (5 g, 10.7 mmol) was dissolved in CH₂Cl₂ (100 mL). The solution was cooled to 0° C., L-prolinamide (1.78 g, 11.7 mmol) and PyBOP® (6.7 g, 12.8 mmol) were added, and the pH adjusted to pH9 with triethylamine. After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (200 mL). The solution was washed with 0.3M KHSO₄ (2×50 mL), sat. NaHCO₃ (2×50 mL), water (2×50 mL) and brine (1×50 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 2% methanol, 98% chloroform) to give a colourless oil identified as (N^(ω)-(tert-butyloxycarbonyl)-N^(ω)-(9-fluorenylmethyloxycarbonyl)-L-lysinyl)-L-prolinamide (4.05 g, 7.2 mmol, 67%).

B. (2S)-1-(N^(α)-(tert-Butyloxycarbonyl)-N^(ω)-(9-fluorenylmethyloxycarbonyl)-L-lysinyl)pyrrolidine-2-carbonitrile

(N^(α)-(tert-Butyloxycarbonyl)-N^(ω)-(9-fluorenylmethyloxycarbonyl)-L-lysinyl)-L-prolinamide (3.95 g, 7.02 mmol) was dissolved in dry THF (100 mL). The solution was cooled to 0° C., triethylamine (1.4 g, 14 mmol) was added followed by the slow addition of trifluoroacetic anhydride (2.97 g, 14.1 mmol). The pH was adjusted to pH9 with triethylamine. After 30 min the reaction mixture was diluted with ethyl acetate (100 mL), washed with water (1×50 mL) and brine (1×50 mL), dried (Na₂SO₄) and evaporated in vacuo to give an orange oil. The residue was purified by flash chromatography on silica gel (eluant: 60% pet ether, 40% ethyl acetate) to give a colourless oil identified as (2S)-1-(N^(α)-(tert-butyloxycarbonyl)-N^(ω)-(9-fluorenylmethyloxycarbonyl)-L-lysinyl)pyrrolidine-2-carbonitrile (3.3 g, 6.11 mmol, 87%).

C. (2S)-1-(N^(α)-(tert-Butyloxycarbonyl)-L-lysinyl)pyrrolidine-2-carbonitrile

(2S)-1-(N^(α)-(tert-Butyloxycarbonyl)-N^(ω)-(9-fluorenylmethyloxycarbonyl)-L-lysinyl)-pyrrolidine-2-carbonitrile (3.1 g, 5.7 mmol) was dissolved in THF (80 mL). Diethylamine (20 mL) was added. After 2 h at room temperature the solvent was removed in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 90% chloroform, 7% methanol, 3% triethylamine) to give a colourless oil identified as (2S)-1-(N^(α)-(tert-butyloxycarbonyl)-L-lysinyl)pyrrolidine-2-carbonitrile (1.63 g, 5.03 mmol, 89%).

D. (2S)-1-(N^(α)-(tert-Butyloxycarbonyl)-N^(ω),N^(ω)-(dicinnamyl)-L-lysinyl)pyrrolidine-2-carbonitrile

(2S)-1-(N^(α)-(tert-Butyloxycarbonyl)-L-lysinyl)pyrrolidine-2-carbonitrile (100 mg, 0.31 mmol) was dissolved in methanol (25 mL). To this solution was added trans-cinnamaldehyde (170 mg, 1.18 mmol). After 30 mins sodium triacetoxyborohydride (330 mg, 1.56 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 2% methanol, 98% chloroform) to give a colourless oil identified as (2S)-1-(N^(α)-(tert-butyloxycarbonyl)-N^(ω),N^(ω)-(dicinnamyl)-L-lysinyl)pyrrolidine-2-carbonitrile (38 mg, 0.068 mmol, 11%). Further elution with 9% methanol, 90% chloroform and 1% acetic acid gave a colourless oil identified as (2S)-1-(N^(α)-(tert-butyloxycarbonyl)-N^(ω)-(cinnamyl)-L-lysinyl)pyrrolidine-2-carbonitrile (32 mg, 0.073 mmol, 12%)

E. (2S)-1-[N^(ω),N^(ω)-(Dicinnamyl)-L-lysinyl]pyrrolidine-2-carbonitrile dihydrochloride

(2S)-1-(N^(α)-(tert-Butyloxycarbonyl)-N^(ω),N^(ω)-(dicinnamyl)-L-lysinyl)pyrrolidine-2-carbonitrile (32 mg, 0.057 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as (2S)-1-[N^(ω),N^(ω)-(dicinnamyl)-L-lysinyl]pyrrolidine-2-carbonitrile dihydrochloride (37 mg, 0.053 mmol, 93%).

[M+H]⁺=457.3

¹H NMR (CD₃OD): δ 1.35-1.55 (2H, m), 1.75-2.00 (2H, m), 2.05-2.23 (6H, m), 3.10-3.29 (4H, m), 3.61-3.68 (2H, m), 4.00-4.03 (4H, m), 4.20-4.30 (1H, m), 4.82-4.93 (1H, m), 6.34-6.39 (2H, m), 6.94 (2H, d, J=5.8 Hz), 7.31-7.37 (6H, m), 7.39-7.53 (4H, m) ppm.

Example 2 (2S)-1-[N^(ω)-(Cinnamyl)-L-lysinyl]pyrrolidine-2-carbonitrile dihydrochloride

A. (2S)-1-[N^(ω)-(Cinnamyl)-L-lysinyl]pyrrolidine-2-carbonitrile dihydrochloride

(2S)-1-(N^(α)-(tert-Butyloxycarbonyl)-N^(ω)-(cinnamyl)-L-lysinyl)pyrrolidine-2-carbonitrile (32 mg, 0.057 mmol). was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as (2S)-1-[N^(ω)-(cinnamyl)-L-lysinyl]pyrrolidine-2-carbonitrile dihydrochloride (37 mg, 0.053 mmol, 93%).

[M+H]⁺=341.5

¹H NMR (CD₃OD): δ 1.29-1.55 (2H, m), 1.72-1.80 (2H, m), 1.90-2.11 (2H, m), 2.16-2.29 (6H, m), 3.02-3.09 (2H, m), 3.65-3.69 (2H, m), 3.78-3.82 (2H, m), 4.23-4.27 (1H, m), 4.81-4.82 (1H, m), 4.91-4.99 (1H, m), 6.21-6.32 (1H, m), 6.86 (1H, d, J=6.1 Hz), 7.26-7.35 (3H, m), 7.37-7.40 (2H, m) ppm.

Example 3 (2S)-1-[N^(ω),N^(ω)-(Dicinnamyl)-L-ornithinyl]pyrrolidine-2-carbonitrile dihydrochloride

A. (2S)-1-(N^(α)-(tert-Butyloxycarbonyl)-L-ornithyl)pyrrolidine-2-carbonitrile

(2S)-1-(N^(α)-(tert-Butyloxycarbonyl)-L-ornithyl)pyrrolidine-2-carbonitrile was prepared by the method described for the lysine derivative in Example 1.

B. (2S)-1-(N^(α)-(tert-Butyloxycarbonyl)-N^(ω),N^(ω)-(dicinnamyl)-L-ornithinyl)pyrrolidine-2-carbonitrile

(2S)-1-(N^(α)-(tert-Butyloxycarbonyl)-L-ornithinyl)pyrrolidine-2-carbonitrile (200 mg, 0.65 mmol) was dissolved in methanol (25 mL). To this solution was added trans-cinnamaldehyde (180 mg, 1.25 mmol). After 30 mins sodium triacetoxyborohydride (343 mg, 1.63 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography (eluant: 2% methanol, 98% chloroform) to give a colourless oil identified as (2S)-1-(N^(α)-(tert-butyloxycarbonyl)-N^(ω),N^(ω)-(dicinnamyl)-L-ornithinyl)-pyrrolidine-2-carbonitrile (77 mg, 0.14 mmol, 22%). Further elution with 9% methanol, 90% chloroform and 1% acetic acid gave a colourless oil identified as (2S)-1-(N^(α)-(tert-butyloxycarbonyl)-N^(ω)-(cinnamyl)-L-ornithinyl)pyrrolidine-2-carbonitrile (78 mg, 0.18 mmol, 28%).

C. (2S)-1-[N^(ω),N^(ω)-(Dicinnamyl)-L-ornithinyl]pyrrolidine-2-carbonitrile dihydrochloride

(2S)-1-(N^(α)-(tert-Butyloxycarbonyl)-N^(ω),N^(ω)-(dicinnamyl)-L-ornithinyl)pyrrolidine-2-carbonitrile (67 mg, 0.12 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as (2S)-1-[N^(ω),N^(ω)-(dicinnamyl)-L-ornithinyl]pyrrolidine-2-carbonitrile dihydrochloride (82 mg, 0.12 mmol, 100%).

[M+H]⁺=443.3

¹H NMR (CD₃OD): δ 1.98-2.12 (4H, m), 2.22-2.29 (4H, m), 3.27-3.31 (4H, m), 3.62-3.67 (2H, m), 3.96 (4H, d, J=7.5 Hz), 4.30-4.40 (1H, m), 4.80-4.83 (1H, m), 6.34-6.41 (2H, m), 6.96 (2H, d, J=15.6 Hz), 7.31-7.39 (6H, m), 7.49-7.53 (4H, m) ppm.

Example 4 (2S)-1-[N^(ω)-(Cinnamyl)-L-ornithinyl]pyrrolidine-2-carbonitrile dihydrochloride

A. (2S)-1-[N^(ω)-(Cinnamyl)-L-ornithinyl]pyrrolidine-2-carbonitrile dihydrochloride

(2S)-1-(N^(α)-(tert-Butyloxycarbonyl)-N^(ω)-(cinnamyl)-L-ornithinyl)pyrrolidine-2-carbonitrile (71 mg, 0.17 mmol). was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as (2S)-1-[N^(ω)-(cinnamyl)-L-ornithinyl]pyrrolidine-2-carbonitrile dihydrochloride (91 mg, 0.16 mmol, 100%).

[M+H]⁺=327.5

¹H NMR (CD₃OD): δ 1.70-1.88 (2H, m), 1.97-2.01 (2H, m), 2.14-2.32 (4H, m), 3.08-3.13 (2H, m), 3.29-3.31 (3H, m), 3.68-3.71 (2H, m), 3.79-3.82 (2H, m), 4.29-4.31 (1H, m), 4.87-4.91 (1H, m), 6.29-6.31 (1H, m), 6.86 (1H, d, J=15.8 Hz), 7.29-7.30 (3H, m), 7.44-7.48 (2H, m) ppm.

Example 5 3-[N^(ω)-N^(ω)-(Dicinnamyl)-L-lysinyl]thiazolidine dihydrochloride

A. 3-[N^(α)-(tert-Butyloxycarbonyl)-N^(ω)-(9-fluorenylmethyloxycarbonyl)-L-lysinyl]-thiazolidine

N^(α)-(tert-Butyloxycarbonyl)-N^(ω)-(9-fluorenylmethyloxycarbonyl)-L-lysine (2.73 g, 6 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 100 mL). To this solution at 0° C. were added 1-hydroxybenzotriazole hydrate (1.53 g, 10 mmol), water-soluble carbodiimide (1.34 g, 7 mmol), thiazolidine (1.28 g, 18 mmol) and N-methylmorpholine (1.0 g, 10 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (100 mL). The solution was washed with 0.3M KHSO₄ (2×25 mL), sat. NaHCO₃ (2×25 mL), water (2×25 mL) and brine (1×25 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 75% ethyl acetate, 25% pet. ether) to give a white solid identified as 3-[N^(α)-(tert-butyloxycarbonyl)-N^(ω)-(9-fluorenylmethyloxycarbonyl)-L-lysinyl]thiazolidine (2.55 g, 4.85 mmol, 81%).

B. 3-[N^(α)-(tert-Butyloxycarbonyl)-L-lysinyl]thiazolidine

3-[N^(α)-(tert-Butyloxycarbonyl)-N^(ω)-(9-fluorenylmethyloxycarbonyl)-L-lysinyl]thiazolidine (1.15 g, 2.13 mmol) was dissolved in acetonitrile (20 mL). Diethylamine (5 mL) was added. After 90 min at room temperature the solvent was removed in vacuo and the residue was purified by flash chromatography on silica gel (eluant: 90% chloroform, 7% methanol, 3% triethylamine) to give a pale yellow oil identified as 3-[N^(α)-(tert-butyloxycarbonyl)-L-lysinyl]thiazolidine (530 mg, 1.67 mmol, 78%).

C. 3-(N^(α)-(tert-Butyloxycarbonyl)-N^(ω),N^(ω)-(dicinnamyl)-L-lysinyl)thiazolidine

3-(N^(α)-(tert-Butyloxycarbonyl)-L-lysinyl)thiazolidine (200 mg, 0.6 mmol) was dissolved in methanol (25 mL). To this solution was added trans-cinnamaldehyde (400 mg, 3.0 mmol). After 30 mins sodium triacetoxyborohydride (534 mg, 2.54 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 2% methanol, 98% chloroform) to give a colourless oil identified as 3-(N^(α)-(tert-butyloxycarbonyl)-N^(ω),N^(ω)-(di-cinnamyl)-L-lysinyl)thiazolidine (139 mg, 0.25 mmol, 40%).

D. 3-[N^(ω),N^(ω)-(Dicinnamyl)-L-lysinyl]thiazolidine dihydrochloride

3(N^(α)-(tert-Butyloxycarbonyl)-N^(ω),N^(ω)-(di-cinnamyl)-L-lysinyl)thiazolidine (139 mg, 0.25 mmol). was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a pale brown solid identified as 3-[N^(ω),N^(ω)-(dicinnamyl)-L-lysinyl]thiazolidine dihydrochloride (127 mg, 0.24 mmol, 96%).

[M+H]⁺=450.2

¹H NMR (CD₃OD): δ 1.49-1.55 (2H,m), 1.89-1.98 (4H, m), 3.01-3.30 (4H, m), 3.4-3.5 (4H, m), 3.7-3.9 (3H, m), 4.0-4.2 (3H, m), 4.2-4.8 (2H, br m), 6.38-6.44 (2H, m), 6.99-6.93 (2H, m), 7.34-7.37 (5H, m), 7.51-7.60 (4H, m) ppm.

Example 6 3-[N^(ω),N^(ω)-(Cinnamyl)-L-lysinyl]thiazolidine dihydrochloride

A. 3-(N^(α)-(tert-Butyloxycarbonyl)-N^(ω),N^(ω)-(cinnamyl)-L-lysinyl)thiazolidine

3-(N^(α)-(tert-Butyloxycarbonyl)-L-lysinyl)thiazolidine (200 mg, 0.6 mmol) was dissolved in methanol (25 mL). To this solution was added trans-cinnamaldehyde (400 mg, 3.0 mmol). After 30 mins sodium triacetoxyborohydride (534 mg, 2.54 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 1% triethylamine, 5% methanol, 94% chloroform) to give a colourless oil identified as 3-(N^(α)-(tert-butyloxycarbonyl)-N^(ω),N^(ω)-(cinnamyl)-L-lysinyl)thiazolidine (215 mg, 0.50 mmol, 83%).

B. 3-[N^(ω),N^(ω)-(Cinnamyl)-L-lysinyl]thiazolidine dihydrochloride

3-(N^(α)-(tert-Butyloxycarbonyl)-N^(ω),N^(ω)-(cinnamyl)-L-lysinyl)thiazolidine (215 mg, 0.5 mmol). was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a pale brown solid identified as 3-[N^(ω),N^(ω)-(cinnamyl)-L-lysinyl]thiazolidine dihydrochloride (160 mg, 0.40 mmol, 79%).

[M+H]⁺=334.4

¹H NMR (CD₃OD): δ 1.28-1.30 (1H, m), 1.51-1.53 (1H, m), 1.79-1.78 (1H, m), 1.93-1.98 (2H, m), 2.9-3.3 (5H, m), 3.6-3.8 (5H, m), 4.30-4.70 (5H, m), 6.2-6.3 (1H, m), 6.85-6.91(1H, m), 7.1-7.7 (5H, m) ppm.

Example 7 1-[N^(ω)-(Cyclohexylmethyl)-L-ornithinyl]pyrolidine dihydrochloride

A. 1-N^(ω)-(Benzyloxycarbonyl)-N^(α)-(tert-butyloxycarbonyl)-L-ornithinyl]pyrrolidine

N^(ω)-(Benzyloxycarbonyl)-N^(α)-(tert-butyloxycarbonyl)-L-ornithine (5.49 g, 15 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 100 mL). To this solution at 0° C. was added 1-hydroxybenzotriazole hydrate (3.37 g, 22 mmol), water-soluble carbodiimide (3.46 g, 18 mmol), pyrrolidine (1.28 g, 18 mmol) and N-methylmorpholine (2.0 g, 20 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (200 mL). The solution was washed with 0.3M KHSO₄ (2×50 mL), sat. NaHCO₃ (2×50 mL), water (2×50 mL) and brine (1×50 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 90% ethyl acetate, 10% pet. ether) to give a colourless oil identified as 1-[N^(ω)-(benzyloxycarbonyl)-N^(α)-(tert-butyloxycarbonyl)-L-ornithyl]pyrrolidine (5.15 g, 12.3 mmol, 82%).

B. 1-[N^(α)-(tert-Butyloxycarbonyl)-L-ornithinyl]pyrrolidine

1-[N^(ω)-(Benzyloxycarbonyl)-N^(α)-(tert-butyloxycarbonyl)-L-ornithinyl]pyrrolidine (2.15 g, 5.13 mmol) was dissolved in methanol (80 mL). This solution was hydrogenated over 10% Pd/C (400 mg). After 2 h the catalyst was filtered off and washed with methanol (50 mL). The combined filtrates were evaporated in vacuo to give an off white solid identified as 1-[N^(α)-(tert-butyloxycarbonyl)-L-ornithinyl]pyrrolidine (1.35 g, 4.74 mmol, 94%).

C. 1-(N^(α)-(tert-Butyloxycarbonyl)-N^(ω)-cyclohexylmethyl)-L-ornithinyl)pyrrolidine

1-[N^(α)-(tert-Butyloxycarbonyl)-L-ornithinyl]pyrrolidine (100 mg, 0.35 mmol) was dissolved in methanol (25 mL). To this solution was added cyclohexanecarboxaldehyde (44 mg, 0.39 mmol). After 30 mins sodium triacetoxyborohydride (148 mg, 0.70 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 1% triethylamine, 5% methanol, 94% chloroform) to give a colourless oil identified as 1-(N^(α)-(tert-Butyloxycarbonyl)-N^(ω)-(cyclohexylmethyl)-L-ornithinyl)pyrrolidine (51 mg, 0.18 mmol, 52%).

D. 1-[N^(ω)-(Cyclohexylmethyl)-L-ornithinyl]pyrrolidine dihydrochloride

1-(N^(α)-(tert-Butyloxycarbonyl)-N^(ω)-(cyclohexylmethyl)-L-ornithinyl)pyrrolidine (215 mg, 0.5 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 1-[N^(ω)-(cyclohexylmethyl)-L-ornithinyl]pyrrolidine dihydrochloride (160 mg, 0.40 mmol, 79%).

[M+H]⁺=282.3

¹H NMR (CD₃OD): δ 0.93-1.24 (3H, m), 1.66-1.81 (15H, m), 2.50-2.70 (2H, m), 2.71-2.88 (2H, m), 3.2-3.48 (6H, m), 4.08 (1H, m), 8.35-8.38 (1H, m), 8.80-8.85 (1H, m) ppm.

Example 8 3-[N^(ω)-Me-N^(ω)-(2-napthylmethyl)-L-lysinyl]thiazolidine dihydrochloride

A. N^(α)-(tert-Butyloxycarbonyl-N^(ω)-benzyl-L-lysine methyl ester

N^(α)-(tert-Butyloxycarbonyl-L-lysine methyl ester (6.1 g, 22.2 mmol) was dissolved in methanol (100 mL). To this solution was added benzaldehyde (1.9 g, 17.5 mmol). After 2 hours sodium triacetoxyborohydride (5.8 g, 27.3 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (200 mL). This solution was washed with sat Na HCO₃ (1×50 mL), water (12×50 mL) and brine (1×50 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 1% acetic acid, 5% methanol, 94% chloroform) to give a colourless oil identified as N^(α)-(tert-butyloxycarbonyl-N^(ω)-benzyl-L-lysine methyl ester (5.2 g, 14.2 mmol, 82%).

B. N^(α)-tert-Butyloxycarbonyl-N^(ω)-benzyl-N^(ω)-methyl-L-lysine methyl ester

N^(α)-tert-Butyloxycarbonyl-N^(ω)-benzyl-L-lysine methyl ester (5.0 g, 14.2 mmol) was dissolved in methanol (100 mL). To this solution was added formaldehyde (37% solution in water, 10 mL). After 2 hours sodium triacetoxyborohydride (3.9 g, 18.4 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (200 mL). This solution was washed with sat. Na HCO₃ (1×50 mL), water (12×50 mL) and brine (1×50 mL), dried (Na₂SO₄) and evaporated in vacuo to give a colourless oil identified as N^(α)-tert-butyloxycarbonyl-N^(ω)-benzyl-N^(ω)-methyl-L-lysine methyl ester (5.2 g, 14.2 mmol, 100%).

C. N^(α)-tert-Butyloxycarbonyl-N^(ω)-methyl-L-lysine methyl ester

N^(α)-tert-Butyloxycarbonyl-N^(ω)-benzyl-N^(ω)-methyl-L-lysine methyl ester (5.0 g, 14.2 mmol) was dissolved in methanol/water (9:1, 100 mL). To this solution was added ammonium formate (1.6, 19.3 mmol) and 10% palladium on charcoal (2 g) . After 3 hours at 60 ° C. the catalyst was filtered off through celite and the residue washed with methanol (50 mL). The combined filtrates were evaporated in vacuo and the residue was taken up in chloroform (200 mL). This solution was washed with sat Na HCO₃ (1×50 mL), water (12×50 mL) and brine (1×50 mL), dried (Na₂SO₄) and evaporated in vacuo to give a colourless oil identified as N^(α)-(tert-butyloxycarbonyl-N^(ω)-methyl-L-lysine methyl ester (3.48 g, 12.5 mmol, 93%).

D. N^(α)-tert-Butyloxycarbonyl-N^(ω)-(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-N^(ω)-methyl-L-lysine methyl ester

N^(α)-tert-Butyloxycarbonyl-N^(ω)-methyl-L-lysine methyl ester (3.1 g, 11.1 mmol) was dissolved in dichloromethane (100 mL). To this solution was added 1,1-dimethyl-2,2,2-trichloroethyl chloroformate (3.0 g, 12.5 mmol) and triethylamine (2.3 g, 23 mmol). After 18 hours at room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (200 mL). This solution was washed with 0.3M KHSO₄ (1×50 mL), sat NaHCO₃ (1×50 mL), water (1×50 mL) and brine (1×50 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil purified by flash chromatography on silica gel (eluant: 30% ethyl acetate, 70% pet. ether) to give colourless oil identified as N^(α)-(tert-butyloxycarbonyl-N^(ω)-(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-N^(ω)-methyl-L-lysine methyl ester (3.28 g, 6.98 mmol, 63%).

E. N⁶⁰ -tert-Butyloxycarbonyl-N^(ω)-(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-N^(ω)-methyl-L-lysine

N^(α)-(tert-Butyloxycarbonyl-N^(ω)-(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-N^(ω)-methyl-L-lysine methyl ester (3.1 g, 6.6 mmol) was dissolved in tetrahydrofuran (100 mL). 1M Lithium hydroxide (7 mL, 7.0 mmol) was added. After 3 hours at room temperature the reaction mixture was diluted with ethyl acetate (150 mL), washed with 1M HCl (1×50 mL), water (1×50 mL) and brine (1×50 mL), dried (Na₂SO₄) and evaporated in vacuo to give colourless oil identified as N^(α)-(tert-butyloxycarbonyl-N^(ω)-(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-N^(ω)-methyl-L-lysine (2.94 g, 6.45 mmol, 98%).

F. 3-(N^(α)-tert-Butyloxycarbonyl-N^(ω)-1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-N^(ω)-methyl-L-lysinyl)thiazoldine

N⁶⁰ -(tert-Butyloxycarbonyl-N^(ω)-(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-N^(ω)-methyl-L-lysine (700 mg, 1.51 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 20 mL). To this solution at 0° C. were added 1-hydroxybenzotriazole hydrate (410 mg, 3.0 mmol), water-soluble carbodiimide (250 mg, 1.3 mmol), thiazolidine (170 mg, 1.9 mmol) and N-methylmorpholine (1.0 g, 10 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (70 mL). The solution was washed with 0.3M KHSO₄ (1×25 mL), sat. NaHCO₃ (1×25 mL), water (1×25 mL) and brine (1×25 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 50% ethyl acetate, 50% pet. ether) to give a white solid identified as 3-(N^(α)-tert-butyloxycarbonyl-N^(ω)-(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-N^(ω)-methyl-L-lysinyl)thiazolidine (758 mg, 1.42 mmol, 94%).

G. 3-(N^(α)-tert-Butyloxycarbonyl-N^(ω)-methyl-L-lysinyl)thiazolidine

3-(N^(α)-tert-Butyloxycarbonyl-N^(ω)-(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-N^(ω)-methyl-L-lysinyl)thiazolidine (730 mg, 1.36 mmol) was dissolved in acetic acid (30 mL). Zinc powder (200 mg) was added. After stirring at room temperature for 18 hours the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). The solution was washed with sat. NaHCO₃ (1×25 mL), water (1×25 mL) and brine (1×25 mL), dried (Na₂SO₄) and evaporated in vacuo to give a colourless oil identified as 3-(N^(α)-tert-butyloxycarbonyl-N^(ω)-methyl-L-lysinyl)thiazolidine (438 mg, 1.32 mmol, 97%).

H. 3-[N^(α)-tert-Butyloxycarbonyl-N^(ω)-methyl-N^(ω)-(2-napthylmethyl)-L-lysinyl]thiazolidine

3-(N^(α)-tert-Butyloxycarbonyl-N^(ω)-methyl-L-lysinyl)thiazolidine (50 mg, 0.15 mmol) was dissolved in 1,2-dichloroethane (20 mL). To this solution was added 2-naphthaldehyde (26 mg, 0.17 mmol). After 2 hours sodium triacetoxyborohydride (36 mg, 0.17 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 4% methanol, 96% chloroform) to give a colourless oil identified as 3-[N^(α)-tert-butyloxycarbonyl-N^(ω)-methyl-N^(ω)-(2-napthylmethyl)-L-lysinyl]thiazolidine (51 mg, 0.11 mmol, 72%).

I. 3-[N^(ω)-Methyl-N^(ω)-(2-napthylmethyl)-L-lysinyl]thiazolidine dihydrochloride

3-[N^(α)-tert-Butyloxycarbonyl-N^(ω)-methyl-N^(ω)-(2-napthylmethyl)-L-lysinyl]thiazolidine (44 mg, 0.093 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a pale brown solid identified as 3-[N^(ω)-methyl-N^(ω)-(2-napthylmethyl)-L-lysinyl]thiazolidine dihydrochloride (37 mg, 0.083 mmol, 89%).

[M+H]⁺=372.2

¹H NMR (CD₃OD): δ 1.50-1.53 (2H,m), 1.91-1.98 (4H,m), 2.82 (3H,s), 3.08-3.19 (4H,m), 3.36-3.75 (5H,m), 4.32-4.47 (2H,m), 4.60-4.71 (2H,m), 7.55-7.59 (2H,m), 7.65-7.68 (1H,m), 7.90-8.00 (3H,m), 8.10-8.12 (1H,m) ppm.

Example 9 3-[N^(ω)-Methyl-N^(ω)-(1-Napthylmethyl)-L-ornithyl]thiazolidine dihydrochloride

A. 3-[N-(tert-Butyloxycarbonyl)-O^(ω)-methyl-L-glutamyl]thiazolidine

N-(tert-Butyloxycarbonyl)-O^(ω)-methyl-L-glutamic acid (6.28 g, 24 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 100 ml). To this solution at 0° C. were added 1-hydroxybenzotriazole hydrate (5.5 g, 36 mmol), water-soluble carbodiimide (5.38 g, 28 mmol), thiazolidine (2.48 g, 28 mmol) and N-methylmorpholine (3.0 g, 30 mmol). The mixture was stirred for 18 h at 0° C. to room temperature then the solvent was removed in vacuo and the residue was taken up in ethyl acetate (150 ml). The solution was washed with 0.3M KHSO₄ (2×30 ml), sat. NaHCO₃ (2×30 ml), water (2×30 ml) and brine (1×30 ml), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 70% ethyl acetate, 30% pet. ether 60-80) to give a brown oil identified as 3-[N-(tert-butyloxycarbonyl)-O^(ω)-methyl-L-glutamyl]thiazolidine (4.0 g, 12 mmol, 50%).

B. 3-[N,N-Di-(tert-butyloxycarbonyl)-O^(ω)-methyl-L-glutamyl]thiazolldine

3-[N-(tert-Butyloxycarbonyl)-O^(ω)-methyl-L-glutamyl]thiazolidine (3.2 g, 9.6 mmol) was dissolved in acetonitrile (20 mL). Di-tert-butyl dicarbonate (3.14 g, 14.4 mmol) and 4-dimethylaminopyridine (235 mg, 1.93 mmol) were added. After 18 hours at room temperature further di-tert-butyl dicarbonate (3.14 g, 14.4 mmol) was added. After a further 3 days at room temperature the solvent was evaporated in vacuo the residue was purified by flash chromatography on silica gel (eluant: 70% ethyl acetate, 30% pet. ether 60-80) to give a colourless oil identified as 3-[N,N-di-(tert-butyloxycarbonyl)-O^(ω)-methyl-L-glutamyl]thiazolidine (2.0 g, 4.63 mmol, 48%).

C. 3-[N,N-Di-(tert-butyloxycarbonyl)-L-glutamyl]thiazolidine

3-N,N-di-(tert-butyloxycarbonyl)-O^(ω)-methyl-L-glutamyl]thiazolidine (950 mg, 2.22 mmol) was dissolved in THF (50 ml). 1M Lithium hydroxide (5.5 ml, 5.5 mmol) was added. The mixture was stirred for 1 hour at room temperature then the solvent was removed in vacuo and the residue was taken up in ethyl acetate (70 ml). The solution was washed with 0.3M KHSO₄ (2×20 ml), water (2×20 ml) and brine (1×20 ml), dried (Na₂SO₄) and evaporated in vacuo to give a colourless oil identified as 3-[N,N-di-(tert-butyloxycarbonyl)-L-glutamyl]thiazolidine (912 mg, 2.2 mmol, 98%).

D. 3-[2-(N,N-Di-(tert-butyloxycarbonyl)amino)-5-hydroxypentanoyl]thiazolidine

3-[N,N-Di-(tert-butyloxycarbonyl)-L-glutamyl]thiazolidine (912 mg, 2.2 mmol) was dissolved in tetrahydrofuran (30 mL). This solution was cooled to −20° C., N-methylmorpholine (300 mg, 2.96 mmol) and isobutyl chloroformate (387 mg, 2.83 mmol) were added. After 20 mins at −20 ° C. the reaction mixture was added to a solution of sodium borohydride (182 mg, 4.8 mmol) in water (5 mL) at 0° C. After 1 hour the reaction mixture was diluted with ethyl acetate (150 mL). This solution was washed with water (1×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a colourless oil identified as 3-[2-(N,N-di-(tert-butyloxycarbonyl)amino-5-hydroxy-pentanoyl]thiazolidine (800 mg, 2.0 mmol, 92%).

E. 3-[2-(N,N-Di-(tert-butyloxycarbonyl)amino-5-oxopentanoyl]thiazolidine

3-[2-N,N-((Di-tert-butyloxycarbonyl)amino)-5-hydroxypentanoyl]thiazolidine (800 mg, 2.0 mmol) was dissolved in dichloromethane (50 mL). Dess-Martin periodinane (933 mg,2.2 mmol) was added. After 1 hour at room temperature the reaction mixture was diluted with ethyl acetate (150 mL). This solution was washed with water (1×20 ml) and brine (1×20 ml), dried (Na₂SO₄) and evaporated in vacuo to give a colourless oil. Purified by flash chromatography on silica gel (eluant: 50% ethyl acetate, 50% pet. ether 60-80) to give a colourless oil identified as 3-[2-(N,N-di-(tert-butyloxycarbonyl)amino-5-oxopentanoyl]thiazolidine (210 mg, 0.52 mmol, 26%).

F. 3-[N,N-Di-(tert-butyloxycarbonyl-N^(ω)-methyl-N^(ω)-(1-napthylmethyl)-L-ornithyl]-thiazolidine

3-[N,N-Di-(tert-butyloxycarbonyl)amino-5-oxopentanoyl]thiazolidine was dissolved in 1,2-dichloroethane (20 mL). To this solution was added N-methyl-1-napthylmethylamine. After 2 hours sodium triacetoxyborohydride was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel to give a colourless oil identified as 3-[N,N-di-(tert-butyloxycarbonyl-N^(ω)-methyl-N^(ω)-(1-napthylmethyl)-L-ornithyl]thiazolidine.

G. 3-[N^(ω)-Methyl-N^(ω)-(1-Napthylmethyl)-L-ornithyl]thiazolidine dihydrochloride

3-[N,N-Di-(tert-butyloxycarbonyl-N^(ω)-methyl-N^(ω)-napthylmethyl)-L-ornithyl]thiazolidine was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a pale brown solid identified as 3-[N^(ω)-Me,N^(ω)-(1-napthylmethyl)-L-ornithyl]thiazolidine dihydrochloride.

Example 10 3,3-Difluoro-1-[N^(ω)-(2-methylbutyl)-L-lysinyl]pyrrolidine dihydrochloride

A. 1-(tert-Butyloxycarbonyl)-3-pyrrolidone

(3R)-1-(tert-Butyloxycarbonyl)-3-hydroxypyrrolidine (980 mg, 5.3 mmol) was dissolved in CH₂Cl₂ (40 ml). Dess-Martin periodinane (2.5 g, 5.8 mmol) was added. The mixture was stirred for 3 hours at room temperature then the solvent was removed in vacuo and the residue was taken up in ethyl acetate (300 ml). The solution was washed with sat. NaHCO₃, water and brine, dried (Na₂SO₄) and evaporated in vacuo to give a colourless oil. The residue was purified by flash chromatography on silica gel (eluant: 20% ethyl acetate, 80% pet. ether 60-80) to give a colourless oil identified as 1-(tert-butyloxycarbonyl)-3-pyrrolidone (842 mg, 4.6 mmol, 87%).

B. 1-(tert-Butyloxycarbonyl)-3,3-difluoropyrrolidine

1-(tert-Butyloxycarbonyl)-3-pyrrolidone (810 mg, 4.4 mmol) was dissolved in CH₂Cl₂ (30 ml). (Diethylamino)sulphur trifluoride (2.2 g, 13.7 mmol) was added to this solution at 0° C. The mixture was stirred for 18 hours at 0° C. to room temperature then carefully poured into sat. NaHCO₃ (100 ml). The mixture was stirred for 15 min then extracted with CH₂Cl₂. The organic extract was washed with water and brine, dried (Na₂SO₄) and evaporated in vacuo to give an orange oil. The residue was purified by flash chromatography (eluant: 10% ethyl acetate, 90% pet. ether 60-80) to give a colourless oil identified as 1-(tert-butyloxycarbonyl)-3,3-difluoropyrrolidine (580 mg, 2.8 mmol, 64%).

C. 3,3-Difluoropyrrolidine hydrochloride

1-(tert-Butyloxycarbonyl)-3,3-difluoropyrrolidine (540 mg, 2.6 mmol) was dissolved in 4M HCl/dioxan (30 ml). The solution was stirred for 1 hour at room temperature then the solvent was removed in vacuo to give an off white solid identified as 3,3-difluoropyrrolidine hydrochloride (370 mg, 2.6 mmol, 100%).

D. 1-[N^(α)-(tert-Butyloxycarbonyl)-N^(ω)-(9-fluorenylmethyloxycarbonyl)-L-lysinyl]-3,3-difluoropyrrolidine

N^(α)-(tert-Butyloxycarbonyl)-N^(ω)-(9-fluorenylmethyloxycarbonyl)-L-lysine (1.14 g, 2.4 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 100 ml). To this solution at 0° C. were added 1-hydroxybenzotriazole hydrate (394 mg, 2.9 mmol), water-soluble carbodiimide (680 mg, 3.4 mmol), 3,3-difluoropyrrolidine hydrochloride (380 mg, 2.43 mmol) and N-methylmorpholine (400 mg, 4 mmol). The mixture was stirred for 18 h at 0° C. to room temperature then the solvent was removed in vacuo and the residue was taken up in ethyl acetate (200 ml). The solution was washed with 0.3M KHSO₄, sat. NaHCO₃, water and brine, dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 65% ethyl acetate, 35% pet. ether 60-80) to give a white solid identified as 1-[N^(α)-(tert-butyloxycarbonyl)-N^(ω)-(9-fluorenylmethyloxycarbonyl)-L-lysinyl]-3,3-difluoropyrrolidine (1.0 g, 1.8 mmol, 75%).

E. 1-[N^(α)-tert-Butyloxycarbonyl)-L-lysinyl]-3,3-difluoropyrrolidine

1-[N^(α)-(tert-Butyloxycarbonyl)-N^(ω)-(9-fluorenylmethyloxycarbonyl)-L-lysinyl]-3,3-difluoro-pyrrolidine (1.01 g, 1.8 mmol) was dissolved in THF (20 ml). Diethylamine (5 ml) was added. The mixture was stirred for 3 hours at room temperature then the solvent was removed in vacuo and the residue was purified by flash chromatography on silica gel (eluant: 90% chloroform, 7% methanol, 3% triethylamine) to give a pale yellow oil identified as 1-[N^(α)-(tert-butyloxycarbonyl)-L-lysinyl]-3,3-difluoropyrrolidine (598 mg, 1.78 mmol, 99%).

F. 1-N^(α)-(tert-Butyloxycarbonyl)-N^(ω)-(2-methylbutyl)-L-lysinyl]-3-difluoro-pyrrolidine

1-[N^(α)-(tert-Butyloxycarbonyl)-L-lysinyl]-3,3-difluoropyrrolidine was dissolved in 1,2-dichloroethane (20 mL). To this solution was added 2-methylbutanal. After 2 hours sodium triacetoxyborohydride was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel to give a colourless oil identified as 1-[N^(α)-(tert-butyloxycarbonyl)-N^(ω)-(2-methylbutyl)-L-lysinyl]-3,3-difluoropyrrolidine.

G. 3,3-Difluoro-1-[N^(ω)-(2-methylbutyl)-L-lysinyl]pyrrolidine dihydrochloride

1-[N^(α)-(tert-Butyloxycarbonyl)-N^(ω)-(2-methylbutyl)-L-lysinyl]-3,3-difluoropyrrolidine was dissolved in 4M HCl/dioxan (20 ml). The mixture was stirred for 1 hour at room temperature then the solvent was removed in vacuo to give a colourless oil identified as 3,3-difluoro-1-[N^(ω)-(2-methylbutyl)-L-lysinyl]pyrrolidine dihydrochloride.

Example 11 1-[N^(ω)-(3-Cyclohexenylmethyl)-L-lysinyl]thiomorpholine dihydrochloride

A. 3-[N^(α)-(tert-Butyloxycarbonyl)-N^(ω)-(9-fluorenylmethyloxycarbonyl)-L-lysinyl]thiomorpholine

N^(α)-(tert-Butyloxycarbonyl)-N^(ω)-(9-fluorenylmethyloxycarbonyl)-L-lysine (2.5 g, 5.34 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 100 mL). To this solution at 0° C. were added 1-hydroxybenzotriazole hydrate (1.44 g, 10.6 mmol), water-soluble carbodiimide (1.35 g, 6.5 mmol), thiomorpholine (710 mg, 6.9 mmol) and N-methylmorpholine (800 mg, 8 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (100 mL). The solution was washed with 0.3M KHSO₄ (2×25 mL), sat. NaHCO₃ (2×25 mL), water (2×25 mL) and brine (1×25 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 75% ethyl acetate, 25% pet. ether) to give a white solid identified as 3-[N^(α)-(tert-butyloxycarbonyl)-N^(ω)-(9-fluorenylmethyloxycarbonyl)-L-lysinyl]thiomorpholine (2.70 g, 4.88 mmol, 91%).

B. 3-[N^(α)-(tert-Butyloxycarbonyl)-L-lysinyl]thiomorpholine

3-[N^(α)-(tert-Butyloxycarbonyl)-N^(ω)-(9-fluorenylmethyloxycarbonyl)-L-lysinyl]thiomorpholine (2.6 g, 4.7 mmol) was dissolved in tetrahydrofuran (20 mL). Diethylamine (5 mL) was added. After 90 min at room temperature the solvent was removed in vacuo and the residue was purified by flash chromatography on silica gel (eluant: 90% chloroform, 7% methanol, 3% triethylamine) to give a pale yellow oil identified as 3-[N^(α)-(tert-butyloxycarbonyl)-L-lysinyl]thiomorpholine (1.2 g, 3.637 mmol, 77%).

C. 3-[N^(α)-(tert-Butyloxycarbonyl)-N^(ω)-(3-cyclohexenylmethyl)-L-lysinyl]-thiomorpholine

3-(N^(α)-(tert-Butyloxycarbonyl)-L-lysinyl)thiomorpholine (150 mg, 0.45 mmol) was dissolved in methanol (25 mL). To this solution was added 3-cyclohexenecarboxaldehyde (400 mg, 0.45 mmol). After 30 mins sodium triacetoxyborohydride (150 mg, 0.71 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 1% acetic acid,9% methanol, 90% chloroform) to give a colourless oil identified as 3-(N^(α)-(tert-butyloxycarbonyl)-N^(ω)-(3-cyclohexenylmethyl)-L-lysinyl)thiomorpholine (66 mg, 0.12 mmol, 26%).

D. 1-[N^(ω)-(3-Cyclohexenylmethyl)-L-lysinyl]thiomorpholine dihydrochloride

3-(N^(α)-(tert-Butyloxycarbonyl)-N^(ω)-(3-cyclohexenylmethyl)-L-lysinyl)thiomorpholine (66 mg, 0.12 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 1-[N^(ω)-(3-cyclohexenylmethyl)-L-lysinyl]thiomorpholine dihydrochloride (62 mg, 0.12 mmol, 100%).

[M+H]⁺=326.2

Example 12 (2S)-1-[N^(ω)-(2-(3′-trifluoromethylanilino)pyridyl-3-carbonyl)-L-ornithyl]thiazoldine dihydrochloride

A. 3-[N⁶⁰ -tert-Butyloxycarbonyl-N^(ω)-(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-L-ornithyl]thiazolidine

N^(α)-(tert-Butyloxycarbonyl-N¹⁰⁷ -(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-L-ornithine (2.5 g, 5.9 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 30 mL). To this solution at 0° C. were added 1-hydroxybenzotriazole hydrate (1.6 g, 11.9 mmol), water-soluble carbodiimide (1.4 g, 7.6 mmol), thiazolidine (650 mg, 7.3 mmol) and N-methylmorpholine (2.0 g, 20 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (70 mL). The solution was washed with 0.3M KHSO₄ (1×25 mL), sat. NaHCO₃ (1×25 mL), water (1×25 mL) and brine (1×25 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 70% ethyl acetate, 30% pet. ether) to give a colourless oil identified as 3-[N^(α)-tert-butyloxycarbonyl-N^(ω)-(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-L-ornithyl]thiazolidine (758 mg, 1.42 mmol, 94%).

B. 3-(N^(α)-tert-Butyloxycarbonyl-L-ornithinyl)thiazolidine

3-[N^(α)-tert-Butyloxycarbonyl-N^(ω)-(1,1-dimethyl-2,2,2-trichloroethoxycarbonyl)-L-ornithyl]thiazolidine (130 mg, 0.26 mmol) was dissolved in acetic acid (30 mL). Zinc powder (100 mg) was added. After stirring at room temperature for 18 hours the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). The solution was washed with sat. NaHCO₃ (1×25 mL), water (1×25 mL) and brine (1×25 mL), dried (Na₂SO₄) and evaporated in vacuo to give a colourless oil identified as 3-(N^(α)-tert-butyloxycarbonyl-L-ornithinyl)thiazolidine (80 mg, 0.26 mmol, 100%).

C. 3-[N^(α)-tert-Butyloxycarbonyl-N^(ω)-(2-(3′-trifluoromethylanilino)pyridyl-3-carbonyl)-L-ornithinyl]thiazolidine

3-(N^(α)-tert-Butyloxycarbonyl-L-ornithinyl)thiazolidine (80 mg, 0.26 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 20 mL). To this solution at 0° C. was added 1-hydroxybenzotriazole hydrate (80 mg, 0.6 mmol), water-soluble carbodiimide (65 mg, 0.32 mmol), niflumic acid (82 mg, 0.29mmol) and N-methylmorpholine (100 mg, 1.0 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (70 mL). The solution was washed with 0.3M KHSO₄ (1×20 mL), sat. NaHCO₃ (1×20 mL), water (1×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 75% ethyl acetate, 25% pet. ether) to give a yellow oil identified as 3-[N^(α)-tert-butyloxycarbonyl-N^(ω)-(2-(3′-trifluoromethylanilino)pyridyl-3-carbonyl)-L-ornithinyl]-thiazolidine (60 mg, 0.12 mmol, 45%).

D. (2S)-1-[N^(ω)-(2-(3′-Trifluoromethylanilino)pyridyl-3-carbonyl)-L-ornithyl]-thiazolidine dihydrochloride

3-[N^(α)-tert-Butyloxycarbonyl-N^(ω)-(2-(3′-trifluoromethylanilino)pyridyl-3-carbonyl)-L-ornithinyl]thiazolidine (54 mg, 0.10 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a pale brown solid identified as (2S)-1-[N^(ω)-(2-(3′-trifluoromethylanilino)pyridyl-3-carbonyl)-L-ornithyl]thiazolidine dihydrochloride (47 mg, 0.10 mmol, 100%).

[M+H]⁺=468.0

¹H NMR (CD₃OD): δ1.77-1.82 (2H, m), 1.84-2.00 (2H, m), 3.03-3.15 (4H, m), 3.41-3.51 (2H, m), 3.65-3.71 (2H, m), 3.80-3.87 (1H, m), 4.46-4.49 (2H, m), 4.65-4.72 (2H, m), 7.06-7.11 (1H, m), 7.61-7.11 (3H, m), 7.95 (1H, s), 8.09 (1H, d, J=4.7 Hz), 8.49 (1H, d, J=4.2 Hz) ppm.

Example 13 3,3-Difluoro-1-[N^(ω)-(2-(3′-chloroanilino)pyridyl-3-carbonyl)-L-ornithyl]pyrrolidine dihydrochloride

A. 1-[N^(α)-(tert-Butyloxycarbonyl)-L-ornithyl]-3,3-difluoropyrrolidine

1-[N^(α)-(tert-Butyloxycarbonyl)-L-ornithyl]-3,3-difluoropyrrolidine was prepared as described for the lysine derivative in Example 9.

B. 3-Chloroanilinonicotinic acid

3-Chloroaniline was dissolved in xylene. 2-Aminonicotinic acid was added. The reaction mixture was heated at 150° C. for 18 hours after which time the reaction mixture was diluted with ethyl acetate giving an off-white solid identified as 3-chloroanilinonicotinic acid.

C. 3,3-Difluoro-[N^(α)-tert-butyloxycarbonyl-N^(ω)-(2-(3′-chloroanilino)pyridyl-3-carbonyl)-L-ornithinyl]pyrrolidine

1-[N^(α)-(tert-Butyloxycarbonyl)-L-ornithyl]-3,3-difluoropyrrolidine was dissolved in CH₂Cl₂/DMF (9:1, 20 mL). To this solution at 0° C. was added 1-hydroxybenzotriazole hydrate, water-soluble carbodiimide, 3-chloroanilinonicotinic acid and N-methylmorpholine.

After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (70 mL). The solution was washed with 0.3M KHSO₄ (1×20 mL), sat. NaHCO₃ (1×20 mL), water (1×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 75% ethyl acetate, 25% pet. ether) to give a yellow oil identified as 3,3-difluoro-[N^(α)-tert-butyloxycarbonyl-N^(ω)-(2-(3′-chloroanilino)pyridyl-3-carbonyl)]-L-ornithinyl)pyrrolidine.

D. 3,3-Difluoro-1-[N^(ω)-(2-(3′-chloroanilino)pyridyl-3-carbonyl)-L-ornithyl]pyrrolidine dihydrochloride

3,3-Difluoro-[N^(α)-tert-butyloxycarbonyl-N^(ω)-(2-(3′-chloroanilino)pyridyl-3-carbonyl)]-L-ornithinyl)pyrrolide was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a pale brown solid identified as 3,3-difluoro-1-[N^(ω)-(2-(3′-chloroanilino)pyridyl-3-carbonyl)-L-ornithyl]pyrrolidine dihydrochloride.

Example 14 3-[N^(ω)-6-Chloro-4-(2′,5′-dichloroanilino)-1,3,5-triazinyl)-L-lysinyl]thiazolidine dihydrochloride

A. 4,6-Dichloro-2-(2′,5′-dichloroanilino)-1,3,5-triazine

Cyanuric chloride (1.844 g, 10 mmol) was dissolved in acetonitrile (20 mL). The solution was cooled to −20° C. A solution of 2,5-dichloroaniline (1.62 g, 10 mmol) and triethylamine (1.0 g, 10 mmol) was slowly added. After 1 hour at −20° C. the solvent was removed in vacuo and the residue was taken up in ethyl acetate (150 mL). The solution was washed with water (1×50 mL) and brine (1×50 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was recrystallised from ethyl acetate/hexane to give an off white solid identified as 4,6-dichloro-2-(2′,5′-dichloroanilino)-1,3,5-triazine (1.86 mg, 6.0 mmol, 60%).

B. 3-[N^(α)-tert-Butyloxycarbonyl-N^(ω)-chloro-4-(2′,5′-dichloroanilino)-1,3,5-triazinyl)-L-lysinyl]thiazolidine

3-(N^(α)-(tert-Butyloxycarbonyl)-L-lysinyl)thiazolidine (800 mg, 2.58 mmol) was dissolved in dichloromethane (30 mL). To this solution was added 4,6-dichloro-2-(2′,5′-dichloroanilino)-1,3,5-triazine (810 mg, 2.6 mmol) and triethylamine (300 mg, 3.0 mmol). After 2 hours at room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (150 mL). This solution was washed with water (2×30 mL) and brine (1×30 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography (eluant: 60% ethyl acetate, 40% pet. ether) to give a white solid identified as 3-[N^(α)-tert-butyloxycarbonyl-N^(ω)-chloro-4-(2′,5′-dichloroanilino)-1,3,5-triazinyl)-L-lysinyl]thiazolidine (1.33 g, 2.23 mmol, 86%).

C. 3-[N^(ω)-6-Chloro-4-(2′,5′-dichloroanilino)-1,3,5-triazinyl)-L-lysinyl]thiazolidine dihydrochloride

3-[N^(α)-tert-Butyloxycarbonyl-N^(ω)-6-chloro-4-(2′,5′-dichloroanilino)-1,3,5-triazinyl)-L-lysinyl]thiazolidine (59 mg, 0.10 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 3-[N^(ω)-6-chloro-4-(2′,5′-dichloroanilino)-1,3,5-triazinyl)-L-lysinyl]thiazolidine dihydrochloride (55 mg, 0.098 mmol, 98%).

[M+H]⁺=492.2, 494.4

¹H NMR (CD₃OD): δ1.46-1.51 (2H,m), 1.65-1.67 (2H,m), 1.80-1.96 (2H, m), 3.05-3.14 (2H,m), 3.38-3.42 (2H,m), 3.55-3.75 (4H,m), 4.31-4.36 (2H,m0, 4.40-4.52 (1H, m), 4.63-4.95 (2H,m), 7.15-7.18 (1H,m), 7.40-7.45 (1H,m), 8.15-8.25 (1H,m) ppm.

Example 15 3-[N^(ω)-4-(2′,5′-Dichloroanilino)-6-hydroxy-1,3,5-triazinyl)-L-lysinyl]thiazolidine bis(trifluoroacetate)

A. 3-[N^(ω)-4-(2′,5′-Dichloroanilino)-6-hydroxy-1,3,5-triazinyl)-L-lysinyl]thiazolidine bis(trifluoroacetate)

3-[N^(α)-tert-Butyloxycarbonyl-N^(ω)-6-chloro-4-(2′,5′-dichloroanilino)-1,3,5-triazinyl)]-L-ornithinyl)thiazolidine (54 mg, 0.09 mmol) was dissolved in trifluoroacetic acid (20 mL) and water (2 mL). After 2 hours at 70° C. the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 3-[N^(ω)-4-(2′,5′-dichloroanilino)-6-hydroxy-1,3,5-triazinyl-L-lysinyl]thiazolidine bis(trifluoroacetate) (63 mg, 0.089 mmol, 97%).

[M+H]⁺=472.1, 474.2

¹H NMR (CD₃OD): δ1.42-1.47 (2H,m), 1.62-1.67 (2H,m), 1.82-1.89 (2H,m), 3.04-3.16 (4H,m), 3.70-3.75 (2H,m), 3.84-3.91 (1H,m), 4.25-4.32 (2H,m), 4.45-4.54 (2H,m), 4.64-4.70 (2H,m), 7.05-7.15 (1H,m), 7.34-7.38 (1H,m), 7.49-7.55 (1H,m), 7.80-7.92 (1H,m) ppm.

Example 16 3-[N^(ω)-4-(2′,5′-Dichloroanilino)-6-methylamino-1,3,5-triazinyl)-L-lysinyl]thiazolidine dihydrochloride

A. 3-[N^(α)-tert-Butyloxycarbonyl-N^(ω)-4-(2′,5′-dichloroanilino)-6-dimethylamino-1,3,5-triazinyl)-L-lysinyl]thiazolidine

3-[N^(α)-tert-Butyloxycarbonyl-N^(ω)-3-chloro-5-(2′,5′-dichloroanilino)-2,4,6-triazinyl)]-L-ornithinyl)thiazolidine (120 mg, 0.20 mmol) was dissolved in 1M dimethylamine in tetrahydrofuran (25 mL). After 18 hours at room temperature the solvent was removed in vacuo and the residue was purified by flash chromatography on silica gel (eluant: 70% ethyl acetate, 30% pet. ether) to give a white solid identified as 3-[N^(α)-tert-butyloxycarbonyl-N^(ω)-4-(2′,5′-dichloroanilino)-6-dimethylamino-1,3,5-triazinyl)-L-lysinyl]thiazolidine (110 mg, 0.18 mmol, 90%).

B. 3-[N^(ω)-4-(2′,5′-Dichloroanilino)-6-dimethylamino-1,3,5-triazinyl)-L-lysinyl]-thiazolidine dihydrochloride

3-[N^(α)-tert-Butyloxycarbonyl-N^(ω)-4-(2′,5′-dichloroanilino)-6-dimethylamino-1,3,5-triazinyl)-L-lysinyl]thiazolidine (110 mg, 0.18 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 3-[N^(ω)-4-(2′,5′-dichloroanilino)-6-dimethylamino-1,3,5-triazinyl)-L-lysinyl]thiazolidine dihydrochloride (105 mg, 0.18 mmol, 100%).

[M+H]⁺=499.1, 501.1

¹H NMR (CD₃OD): δ1.52-1.55 (2H,m), 1.69-1.71 (2H,m), 1.90-1.98 (2H,m), 3.13-3.22 (8H,m), 3.42-3.62 (2H,m), 3.65-3.69 (4H,m), 4.37-4.39 (2H,m), 4.46-4.49 (1H,m), 4.57-4.77 (2H,m), 7.20-7.22 (1H,m), 7.45-7.50 (1H,m), 8.09-8.12 (1H,m) ppm.

The following compounds were prepared by analogous methods. TABLE 1

Example No n X 17 3

18 19 3 4

20 21 3 4

22 23 3 4

24 25 3 4

26 27 3 4

TABLE 2

Example No n X 28 2

29 30 31 2 3 4

32 33 34 2 3 4

35 36 37 2 3 4

38 39 40 2 3 4

41 2

42 43 45 2 3 4

46 47 48 2 3 4

49 2

50 51 52 2 3 4

TABLE 3

Ex No a b X R³ R⁴ 53 1 3 S H

54 1 4 H 55 1 3 CH₂ H 56 1 4 H 57 1 3 CF₂ H 58 1 4 H 59 1 4 S CH₃ 60 1 4 CH(CH₃)₂ 61 1 4 CH₂ CH₃ 62 1 4 CH(CH₃)₂ 63 1 3 S CH(CH₃)₂ 64 1 3 CH₂ CH(CH₃)₂ 65 2 3 S H 66 2 4 H 67 2 3 CH₂ H 68 2 4 H 69 1 3 S H

70 1 4 H 71 1 3 CH₂ H 72 1 4 H 73 1 3 CF₂ H 74 1 4 H 75 1 4 S CH₃ 76 1 4 CH(CH₃)₂ 77 1 4 CH₂ CH₃ 78 1 4 CH(CH₃)₂ 79 1 3 S CH(CH₃)₂ 80 1 3 CH₂ CH(CH₃)₂ 81 2 3 S H 82 2 4 H 83 2 3 CH₂ H 84 2 4 H 85 1 3 S H

86 1 4 H 87 1 3 CH₂ H 88 1 4 H 89 1 3 CF₂ H 90 1 4 H 91 1 4 S CH₃ 92 1 4 CH(CH₃)₂ 93 1 4 CH₂ CH₃ 94 1 4 CH(CH₃)₂ 95 1 3 S CH(CH₃)₂ 96 1 3 CH₂ CH(CH₃)₂ 97 2 3 S H 98 2 4 H 99 2 3 CH₂ H 100 2 4 H 101 1 3 S H

102 1 4 H 103 1 3 CH₂ H 104 1 4 H 105 1 3 CF₂ H 106 1 4 S CH₃ 107 1 4 CH(CH₃)₂ 108 1 4 CH₂ CH₃ 109 1 4 CH(CH₃)₂ 110 1 3 S CH(CH₃)₂ 111 1 3 CH₂ CH(CH₃)₂ 112 2 3 S H 113 2 4 H 114 2 3 CH₂ H 115 2 4 H 116 1 3 S H

117 1 4 H 118 1 3 CH₂ H 119 1 4 H 120 1 3 CF₂ H 121 1 4 H 122 1 4 S CH₃ 123 1 4 CH(CH₃)₂ 124 1 4 CH₂ CH₃ 125 1 4 CH(CH₃)₂ 126 1 3 S CH(CH₃)₂ 127 1 3 CH₂ CH(CH₃)₂ 128 2 3 S H 129 2 4 H 130 2 3 CH₂ H 131 2 4 H 132 1 3 S H

133 1 4 H 134 1 3 CH₂ H 135 1 4 H 136 1 3 CF₂ H 137 1 4 H 138 1 4 S CH₃ 139 1 4 CH(CH₃)₂ 140 1 4 CH₂ CH₃ 141 1 4 CH(CH₃)₂ 142 1 3 S CH(CH₃)₂ 143 1 3 CH₂ CH(CH₃)₂ 144 2 3 S H 145 2 4 H 146 2 3 CH₂ H 147 2 4 H 148 1 3 S

149 1 4 150 1 4 CH₂ 151 1 3 CF₂ 152 1 4 153 1 4 S CH₃ 154 1 4 CH(CH₃)₂ 155 1 4 CH₂ CH₃ 156 1 4 CH(CH₃)₂ 157 1 3 S CH(CH₃)₂ 158 1 3 CH₂ CH(CH₃)₂ 159 2 3 S H 160 2 4 H 161 2 3 CH₂ H 162 2 4 H 163 1 3 S H

164 1 4 H 165 1 3 CH₂ H 166 1 4 H 167 1 3 CF₂ H 168 1 4 H 169 1 4 S CH₃ 170 1 4 CH(CH₃)₂ 171 1 4 CH₂ CH₃ 172 1 4 CH(CH₃)₂ 173 1 3 S CH(CH₃)₂ 174 1 3 CH₂ CH(CH₃)₂ 175 2 3 S H 176 2 4 H 177 2 3 CH₂ H 178 2 4 H 179 1 3 S H

180 1 4 H 181 1 3 CH₂ H 182 1 4 H 183 1 3 CF₂ H 184 1 4 H 185 1 4 S CH₃ 186 1 4 CH(CH₃)₂ 187 1 4 CH₂ CH₃ 188 1 4 CH(CH₃)₂ 189 1 3 S CH(CH₃)₂ 190 1 3 CH₂ CH(CH₃)₂ 191 2 3 S H 192 2 4 H 193 2 3 CH₂ H 194 2 4 H 195 1 3 S H

196 1 4 H 197 1 3 CH₂ H 198 1 4 H 199 1 3 CF₂ H 200 1 4 H 201 1 4 S CH₃ 202 1 4 CH(CH₃)₂ 203 1 4 CH₂ CH₃ 204 1 4 CH(CH₃)₂ 205 1 3 S CH(CH₃)₂ 206 1 3 CH₂ CH(CH₃)₂ 207 2 3 S H 208 2 4 H 209 2 3 CH₂ H 210 2 4 H 211 1 3 S H

212 1 4 H 213 1 3 CH₂ H 214 1 4 H 215 1 3 CF₂ H 216 1 4 H 217 1 4 S CH₃ 218 1 4 CH(CH₃)₂ 219 1 4 CH₂ CH₃ 220 1 4 CH(CH₃)₂ 221 1 3 S CH(CH₃)₂ 222 1 3 CH₂ CH(CH₃)₂ 223 2 3 S H 224 2 3 CH₂ H 225 2 4 H 226 1 3 S H

227 1 4 H 228 1 3 CH₂ H 229 1 4 H 230 1 3 CF₂ H 231 1 4 H 232 1 4 S CH₃ 233 1 4 CH(CH₃)₂ 234 1 4 CH₂ CH₃ 235 1 4 CH(CH₃)₂ 236 1 3 S CH(CH₃)₂ 237 1 3 CH₂ CH(CH₃)₂ 238 2 3 S H 239 2 4 H 240 2 3 CH₂ H 241 2 4 H 242 1 3 S H

243 1 4 H 244 1 3 CH₂ H 245 1 4 H 246 1 3 CF₂ H 247 1 4 H 248 1 4 S CH₃ 249 1 4 CH(CH₃)₂ 250 1 4 CH₂ CH₃ 251 1 4 CH(CH₃)₂ 252 1 3 S CH(CH₃)₂ 253 1 3 CH₂ CH(CH₃)₂ 254 2 3 S H 255 2 4 H 256 2 3 CH₂ H 257 2 4 H 258 1 3 S H

259 1 4 H 260 1 3 CH₂ H 261 1 4 H 262 1 3 CF₂ H 263 1 4 H 264 1 4 S CH₃ 265 1 4 CH(CH₃)₂ 266 1 4 CH₂ CH₃ 267 1 4 CH(CH₃)₂ 268 1 3 S CH(CH₃)₂ 269 1 3 CH₂ CH(CH₃)₂ 270 2 3 S H 271 2 4 H 272 2 3 CH₂ H 273 2 4 H 274 1 3 S H

275 1 4 H 276 1 3 CH₂ H 277 1 4 H 278 1 3 CF₂ H 279 1 4 H 280 1 4 S CH₃ 281 1 4 CH(CH₃)₂ 282 1 4 CH₂ CH₃ 283 1 4 CH(CH₃)₂ 284 1 3 S CH(CH₃)₂ 285 1 3 CH₂ CH(CH₃)₂ 286 2 3 S H 287 2 4 H 288 2 3 CH₂ H 289 2 4 H 290 1 3 S H

291 1 4 H 292 1 3 CH₂ H 293 1 4 H 294 1 3 CF₂ H 295 1 4 H 296 1 4 S CH₃ 297 1 4 CH(CH₃)₂ 298 1 4 CH₂ CH₃ 299 1 4 CH(CH₃)₂ 300 1 3 S CH(CH₃)₂ 301 1 3 CH₂ CH(CH₃)₂ 302 2 3 S H 303 2 4 H 304 2 3 CH₂ H 305 2 4 H 306 1 3 S H

307 1 4 H 308 1 3 CH₂ H 309 1 4 H 310 1 3 CF₂ H 311 1 4 H 312 1 4 S CH₃ 313 1 4 CH(CH₃)₂ 314 1 4 CH₂ CH₃ 315 1 4 CH(CH₃)₂ 316 1 3 S CH(CH₃)₂ 317 1 3 CH₂ CH(CH₃)₂ 318 2 3 S H 319 2 4 H 320 2 3 CH₂ H 321 2 4 H 322 1 3 S H

323 1 4 H 324 1 3 CH₂ H 325 1 4 H 326 1 3 CF₂ H 327 1 4 H 328 1 4 S CH₃ 329 1 4 CH(CH₃)₂ 330 1 4 CH₂ CH₃ 331 1 4 CH(CH₃)₂ 332 1 3 S CH(CH₃)₂ 333 1 3 CH₂ CH(CH₃)₂ 334 2 3 S H 335 2 4 H 336 2 3 CH₂ H 337 2 4 H 338 1 3 S H

339 1 4 H 340 1 3 CH₂ H 341 1 4 H 342 1 3 CF₂ H 343 1 4 H 344 1 4 S CH₃ 345 1 4 CH(CH₃)₂ 346 1 4 CH₂ CH₃ 347 1 4 CH(CH₃)₂ 348 1 3 S CH(CH₃)₂ 349 1 3 CH₂ CH(CH₃)₂ 350 2 3 S H 351 2 4 H 352 2 3 CH₂ H 353 2 4 H 354 1 3 S H

355 1 4 H 356 1 3 CH₂ H 357 1 4 H 358 1 3 CF₂ H 359 1 4 H 360 1 4 S CH₃ 361 1 4 CH(CH₃)₂ 362 1 4 CH₂ CH₃ 363 1 4 CH(CH₃)₂ 364 1 3 S CH(CH₃)₂ 365 1 3 CH₂ CH(CH₃)₂ 366 2 3 S H 367 2 4 H 368 2 3 CH₂ H 369 2 4 H 370 1 3 S H

371 1 4 H 372 1 3 CH₂ H 373 1 4 H 374 1 3 CF₂ H 375 1 4 H 376 1 4 S CH₃ 377 1 4 CH(CH₃)₂ 378 1 4 CH₂ CH₃ 379 1 4 CH(CH₃)₂ 380 1 3 S CH(CH₃)₂ 381 1 3 CH₂ CH(CH₃)₂ 382 2 3 S H 383 2 4 H 384 2 3 CH₂ H 385 2 4 H 386 1 3 S H

387 1 4 H 388 1 3 CH₂ H 389 1 4 H 390 1 3 CF₂ H 391 1 4 H 392 1 4 S CH₃ 393 1 4 CH(CH₃)₂ 394 1 4 CH₂ CH₃ 395 1 4 CH(CH₃)₂ 396 1 3 S CH(CH₃)₂ 397 1 3 CH₂ CH(CH₃)₂ 398 2 3 S H 399 2 4 H 400 2 3 CH₂ H 401 2 4 H 402 1 3 S H

403 1 4 H 404 1 3 CH₂ H 405 1 4 H 406 1 3 CF₂ H 407 1 4 H 408 1 4 S CH₃ 409 1 4 CH(CH₃)₂ 410 1 4 CH₂ CH₃ 411 1 4 CH(CH₃)₂ 412 1 3 S CH(CH₃)₂ 413 1 3 CH₂ CH(CH₃)₂ 414 2 3 S H 415 2 4 H 416 2 3 CH₂ H 417 2 4 H 418 1 3 S H

419 1 4 H 420 1 3 CH₂ H 421 1 4 H 422 1 3 CF₂ H 423 1 4 H 424 1 4 S CH₃ 425 1 4 CH(CH₃)₂ 426 1 4 CH₂ CH₃ 427 1 4 CH(CH₃)₂ 428 1 3 S CH(CH₃)₂ 429 1 3 CH₂ CH(CH₃)₂ 450 2 3 S H 451 2 4 H 452 2 3 CH₂ H 453 2 4 H 454 1 3 S H

455 1 4 H 456 1 3 CH₂ H 457 1 4 H 458 1 3 CF₂ H 459 1 4 H 460 1 4 S CH₃ 461 1 4 CH(CH₃)₂ 462 1 4 CH₂ CH₃ 463 1 4 CH(CH₃)₂ 464 1 3 S CH(CH₃)₂ 465 1 3 CH₂ CH(CH₃)₂ 466 2 3 S H 467 2 4 H 468 2 3 CH₂ H 469 2 4 H 470 1 3 S H

471 1 4 H 472 1 3 CH₂ H 473 1 4 H 474 1 3 CF₂ H 475 I 4 H 476 1 4 S CH₃ 477 1 4 CH(CH₃)₂ 478 1 4 CH₂ CH₃ 479 1 4 CH(CH₃)₂ 480 1 3 S CH(CH₃)₂ 481 1 3 CH₂ CH(CH₃)₂ 482 2 3 S H 483 2 4 H 484 2 3 CH₂ H 485 2 4 H 486 1 3 S H

487 1 4 H 488 1 3 CH₂ H 489 1 4 H 490 1 3 CF₂ H 491 1 4 H 492 1 4 S CH₃ 493 1 4 CH(CH₃)₂ 494 1 4 CH₂ CH₃ 495 1 4 CH(CH₃)₂ 496 1 3 S CH(CH₃)₂ 497 1 3 CH₂ CH(CH₃)₂ 498 2 3 S H 499 2 4 H 500 2 3 CH₂ H 501 2 4 H 502 1 3 S H

503 1 4 H 504 1 3 CH₂ H 505 1 4 H 506 1 3 CF₂ H 507 1 4 H 508 1 4 S CH₃ 509 1 4 CH(CH₃)₂ 510 1 4 CH₂ CH₃ 511 1 4 CH(CH₃)₂ 512 1 3 S CH(CH₃)₂ 513 1 3 CH₂ CH(CH₃)₂ 514 2 3 S H 515 2 4 H 516 2 3 CH₂ H 517 2 4 H 518 1 3 S H

519 1 4 H 520 1 3 CH₂ H 521 1 4 H 522 1 3 CF₂ H 523 1 4 H 524 1 4 S CH₃ 525 1 4 CH(CH₃)₂ 526 1 4 CH₂ CH₃ 527 1 4 CH(CH₃)₂ 528 1 3 S CH(CH₃)₂ 529 1 3 CH₂ CH(CH₃)₂ 530 2 3 S H 531 2 4 H 532 2 3 CH₂ H 533 2 4 H 534 1 3 S H

535 1 4 H 536 1 3 CH₂ H 537 1 4 H 538 1 3 CF₂ H 539 1 4 H 540 1 4 S CH₃ 541 1 4 CH(CH₃)₂ 542 1 4 CH₂ CH₃ 543 1 4 CH(CH₃)₂ 544 1 3 S CH(CH₃)₂ 545 1 3 CH₂ CH(CH₃)₂ 546 2 3 S H 547 2 4 H 548 2 3 CH₂ H 549 2 4 H 550 1 3 S H

551 1 4 H 552 1 3 CH₂ H 553 1 4 H 554 1 3 CF₂ H 555 1 4 H 556 1 4 S CH₃ 557 1 4 CH(CH₃)₂ 558 1 4 CH₂ CH₃ 559 1 4 CH(CH₃)₂ 560 1 3 S CH(CH₃)₂ 561 1 3 CH₂ CH(CH₃)₂ 562 2 3 S H 563 2 4 H 564 2 3 CH₂ H 565 2 4 H 566 1 3 S H

567 1 4 H 568 1 3 CH₂ H 569 1 4 H 570 1 3 CF₂ H 571 1 4 H 572 1 4 S CH₃ 573 1 4 CH(CH₃)₂ 574 1 4 CH₂ CH₃ 575 1 4 CH(CH₃)₂ 576 1 3 S CH(CH₃)₂ 577 1 3 CH₂ CH(CH₃)₂ 578 2 3 S H 579 2 4 H 580 2 3 CH₂ H 581 2 4 H 582 1 3 S H

583 1 4 H 584 1 3 CH₂ H 585 1 4 H 586 1 3 CF₂ H 587 1 4 H 588 1 4 S CH₃ 589 1 4 CH(CH₃)₂ 590 1 4 CH₂ CH₃ 591 1 4 CH(CH₃)₂ 592 1 3 S CH(CH₃)₂ 593 1 3 CH₂ CH(CH₃)₂ 594 2 3 S H 595 2 4 H 596 2 3 CH₂ H 597 2 4 H 598 1 3 S H

599 1 4 H 600 1 3 CH₂ H 601 1 4 H 602 1 3 CF₂ H 603 1 4 H 604 1 4 S CH₃ 605 1 4 CH(CH₃)₂ 606 1 4 CH₂ CH₃ 607 1 4 CH(CH₃)₂ 608 1 3 S CH(CH₃)₂ 609 1 3 CH₂ CH(CH₃)₂ 610 2 3 S H 611 2 4 H 612 2 3 CH₂ H 613 2 4 H 614 1 3 S H

615 1 4 H 616 1 3 CH₂ H 617 1 4 H 618 1 3 CF₂ H 619 1 4 H 620 1 4 S CH₃ 621 1 4 CH(CH₃)₂ 622 1 4 CH₂ CH₃ 623 1 4 CH(CH₃)₂ 624 1 3 S CH(CH₃)₂ 625 1 3 CH₂ CH(CH₃)₂ 626 2 3 S H 627 2 4 H 628 2 3 CH₂ H 629 2 4 H 630 1 3 S H

631 1 4 H 632 1 3 CH₂ H 633 1 4 H 634 1 3 CF₂ H 635 1 4 H 636 1 4 S CH₃ 637 1 4 CH(CH₃)₂ 638 1 4 CH₂ CH₃ 639 1 4 CH(CH₃)₂ 640 1 3 S CH(CH₃)₂ 641 1 3 CH₂ CH(CH₃)₂ 642 2 3 S H 643 2 4 H 644 2 3 CH₂ H 645 2 4 H 646 1 3 S H

647 1 4 H 648 1 3 CH₂ H 649 1 4 H 650 1 3 CF₂ H 651 1 4 H 652 1 4 S CH(CH₃)₂ 653 1 4 CH₂ CH₃ 654 1 4 CH(CH₃)₂ 655 1 3 S CH(CH₃)₂ 656 1 3 CH₂ CH(CH₃)₂ 657 2 3 S H 658 2 4 H 659 2 3 CH₂ H 660 2 4 H 661 1 3 S H

662 1 4 H 663 1 3 CH₂ H 664 1 4 H 665 1 3 CF₂ H 666 1 4 H 667 1 4 S CH₃ 668 1 4 CH(CH₃)₂ 669 1 4 CH₂ CH₃ 670 1 4 CH(CH₃)₂ 671 1 3 S CH(CH₃)₂ 672 1 3 CH₂ CH(CH₃)₂ 673 2 3 S H 674 2 4 H 675 2 3 CH₂ H 676 2 4 H 677 1 3 S H

678 1 4 CH₂ H 679 1 3 CF₂ H 680 1 4 H 681 1 4 S CH₃ 682 1 4 CH(CH₃)₂ 683 1 4 CH₂ CH₃ 684 1 4 CH(CH₃)₂ 685 1 3 S CH(CH₃)₂ 686 1 3 CH₂ CH(CH₃)₂ 687 2 3 S H 688 2 4 CH₂ H 689 1 3 S H

690 1 4 H 691 1 3 CH₂ H 692 1 4 H 693 1 3 CF₂ H 694 1 4 H 695 1 4 S CH₃ 696 1 4 CH(CH₃)₂ 697 1 4 CH₂ CH₃ 698 1 4 CH(CH₃)₂ 699 1 3 S CH(CH₃)₂ 700 1 3 CH₂ CH(CH₃)₂ 701 2 3 S H 702 2 4 H 703 2 3 CH₂ H 704 2 4 H 705 1 3 S H

706 1 4 H 707 1 3 CH₂ H 708 1 4 H 709 1 3 CF₂ H 710 1 4 H 711 1 4 S CH₃ 712 1 4 CH(CH₃)₂ 713 1 4 CH₂ CH₃ 714 1 4 CH(CH₃)₂ 715 1 3 S CH(CH₃)₂ 716 1 3 CH₂ CH(CH₃)₂ 717 2 3 S H 718 2 4 H 719 2 3 CH₂ H 720 2 4 H 721 1 3 S H

722 1 4 H 723 1 3 CH₂ H 724 1 4 H 725 1 3 CF₂ H 726 1 4 H 727 1 4 S CH₃ 728 1 4 CH(CH₃)₂ 729 1 4 CH₂ CH₃ 730 1 4 CH(CH₃)₂ 731 1 3 S CH(CH₃)₂ 732 1 3 CH₂ CH(CH₃)₂ 733 2 3 S H 734 2 4 H 735 2 3 CH₂ H 736 2 4 H 737 1 3 S H

738 1 3 CH₂ H 739 1 4 H 740 1 3 CH₂ H 741 1 4 H 742 1 4 S CH₃ 743 1 4 CH(CH₃)₂ 744 1 4 CH₂ CH₃ 745 1 4 CH(CH₃)₂ 746 1 3 S CH(CH₃)₂ 747 1 3 CH₂ CH(CH₃)₂ 748 2 3 S H 749 2 4 H 750 2 3 CH₂ H 751 2 4 H 752 1 3 S H

753 1 4 H 754 1 3 CH₂ H 755 1 4 H 756 1 3 CF₂ H 757 1 4 H 758 1 4 S CH₃ 759 1 4 CH(CH₃)₂ 760 1 4 CH₂ CH₃ 761 1 4 CH(CH₃)₂ 762 1 3 S CH(CH₃)₂ 763 1 3 CH₂ CH(CH₃)₂ 764 2 3 S H 765 2 4 H 766 2 3 CH₂ H 767 2 4 H 768 1 3 S H

769 1 4 H 770 1 3 CH₂ H 771 1 4 H 772 1 3 CF₂ H 773 1 4 H 774 1 4 S CH₃ 775 1 4 CH(CH₃)₂ 776 1 4 CH₂ CH₃ 777 1 4 CH(CH₃)₂ 778 1 3 S CH(CH₃)₂ 779 1 3 CH₂ CH(CH₃)₂ 780 2 3 S H 781 2 4 H 782 2 3 CH₂ H 783 2 4 H 784 1 3 S H 785 1 4 H 786 1 3 CH₂ H 787 1 4 H 788 1 3 CF₂ H 789 1 4 H 790 1 4 S CH₃ 791 1 4 CH(CH₃)₂ 792 1 4 CH₂ CH₃ 793 1 4 CH(CH₃)₂ 794 1 3 S CH(CH₃)₂ 795 1 3 CH₂ CH(CH₃)₂ 796 2 3 S H 797 2 4 H 798 2 3 CH₂ H 799 2 4 H

TABLE 4

Example No X R 800 801 S CH₂

802 803 S CH₂

804 805 S CH₂

806 807 S CH₂

808 809 S CH₂

810 811 S CH₂

812 813 S CH₂

814 815 S CH₂

816 817 S CH₂

818 819 S CH₂

820 821 S CH₂

822 823 S CH₂

824 825 S CH₂

826 827 S CH₂

828 829 CH₂

830 831 S CH₂

832 833 S CH₂

834 835 S CH₂

836 837 S CH₂

838 839 S CH₂

841 842 S CH₂

843 844 S CH₂

845 846 S CH₂

847 S CH₂

848 849 S CH₂

850 851 S CH₂

852 853 S CH₂

854 855 S CH₂

856 857 S CH₂

858 859 S CH₂

860 861 S CH₂

862 863 S CH₂

864 865 S CH₂

866 867 S CH₂

868 869 S CH₂

870 871 S CH₂

872 873 S CH₂

874 875 S CH₂

876 877 S CH₂

TABLE 5

Example No n X R 878 879 880 881 3 4 3 4 S CH₂

882 883 884 885 3 4 3 4 S CH₂

886 887 888 889 3 4 3 4 S CH₂

890 891 892 893 3 4 3 4 S CH₂

894 895 896 897 3 4 3 4 S CH₂

898 899 900 901 3 4 3 4 S CH₂

902 903 904 905 3 4 3 4 S CH₂

906 907 908 909 3 4 3 4 S CH₂

910 911 912 913 3 4 3 4 S CH₂

914 915 916 917 3 4 3 4 S CH₂

918 919 920 0 3 4 3 4 S CH₂

921 922 923 924 3 4 3 4 S CH₂

925 926 927 928 3 4 3 4 S CH₂

929 3 S Me 930 4 931 3 CH₂ 932 4 933 934 935 936 3 4 3 4 S CH₂

937 938 939 940 3 4 3 4 S CH₂

941 942 943 944 3 4 3 4 S CH₂

945 946 3 4 S CH₂

947 948 949 950 3 4 3 4 S CH₂

951 952 953 954 3 4 3 4 S CH₂

955 956 957 958 3 4 3 4 S CH₂

959 960 961 962 3 4 3 4 S CH₂

963 964 965 966 3 4 3 4 S CH₂

967 968 969 970 3 4 3 4 S CH₂

971 972 973 974 3 4 3 4 S CH₂

975 976 977 978 4 3 4 S CH₂

979 3 S MeS 980 4 981 3 CH₂ 982 4 983 3 S MeO 984 4 985 3 CH₂ 986 4

TABLE 6

Example No n X R 987 988 989 990 3 4 3 4 S CH₂

991 992 993 994 4 3 4 S CH₂

995 996 997 998 3 4 3 4 S CH₂

999 1000 1001 1002 3 4 3 4 S CH₂

1003 1004 1005 1006 3 4 3 4 S CH₂

1007 1008 1009 1010 3 4 3 4 S CH₂

1011 1012 1013 1014 3 4 3 4 S CH₂

1015 1016 1017 1018 3 4 3 4 S CH₂

1019 1020 1021 1022 3 4 3 4 S CH₂

1023 1024 1025 1026 3 4 3 4 S CH₂

1027 1028 1029 1030 3 4 3 4 S CH₂

1031 1032 1033 1034 3 4 3 4 S CH₂

1035 1036 1037 1038 3 4 3 4 S CH₂

1039 3 S Me 1040 4 1041 3 CH₂ 1042 4 1044 1045 1046 1047 3 4 3 4 S CH₂

1048 1049 1050 1051 3 4 3 4 S CH₂

1052 1053 1054 1055 3 4 3 4 S CH₂

1056 1057 3 4 S CH₂

1058 1059 1060 1061 3 4 3 4 S CH₂

1062 1063 1064 1065 3 4 3 4 S CH₂

1066 1067 1068 1069 3 4 3 4 S CH₂

1070 1071 1072 1073 3 4 3 4 S CH₂

1074 1075 1076 1077 3 4 3 4 S CH₂

1078 1079 1080 1081 3 4 3 4 S CH₂

1082 1083 1084 1085 3 4 3 4 S CH₂

1086 1087 1088 1089 3 4 3 4 S CH₂

1090 3 S MeS 1091 4 1092 3 CH₂ 1093 4 1094 3 S MeO 1095 4 1096 3 CH₂ 1097 4

TABLE 7

Example No n X R 1098 1099 1100 1101 3 4 3 4 S CH₂

1102 1103 1104 1105 3 4 3 4 S CH₂

1106 1107 1108 1109 3 4 3 4 S CH₂

1110 1111 1112 1113 3 4 3 4 S CH₂

1114 1115 1116 1117 3 4 3 4 S CH₂

1118 1119 1120 1121 3 4 3 4 S CH₂

1122 1123 1124 1125 3 4 3 4 S CH₂

1125a 1126 1127 1128 3 4 3 4 S CH₂

1129 1130 1131 1132 3 4 3 4 S CH₂

1133 1134 1135 1136 3 4 3 4 S CH₂

1137 1138 1139 1140 3 4 3 4 S CH₂

1141 1142 1143 1144 3 4 3 4 S CH₂

1145 1146 1147 1148 3 4 3 4 S CH₂

1149 1150 1151 1152 3 4 3 4 S CH₂

1153 1154 1155 1156 3 4 3 4 S CH₂

1157 1158 3 4 S CH₂

1159 1160 1161 1162 3 4 3 4 S CH₂

1163 1164 1165 1166 3 4 3 4 S CH₂

1167 1168 1169 1170 3 4 3 4 S CH₂

1171 1172 1173 1174 3 4 3 4 S CH₂

1175 1176 1177 1178 3 4 3 4 S CH₂

1179 1180 1181 1182 3 4 3 4 S CH₂

1183 1184 1185 1186 3 4 3 4 S CH₂

TABLE 8

Example No n X R 1187 1188 1189 1190 3 4 3 4 S CH₂

1191 1192 1193 1194 3 4 3 4 S CH₂

1195 1196 1197 1198 3 4 3 4 S CH₂

1199 1200 1201 1202 3 4 3 4 S CH₂

1203 1204 1205 1206 3 4 3 4 S CH₂

1207 1208 1209 1210 3 4 3 4 S CH₂

1211 1212 1213 1214 3 4 3 4 S CH₂

1215 1216 1217 1218 3 4 3 4 S CH₂

1219 1220 1221 1222 3 4 3 4 S CH₂

1223 1224 1225 1226 3 4 3 4 S CH₂

1227 1228 1229 1230 3 4 3 4 S CH₂

1231 1232 1233 1235 3 3 4 S CH₂

1235 1236 1237 1238 3 4 3 4 S CH₂

1239 1240 1241 1242 3 4 3 4 S CH₂

1243 1244 1245 1246 3 4 3 4 S CH₂

1247 1248 3 4 S CH₂

1249 1250 1251 1252 3 4 3 4 S CH₂

1253 1254 1255 1256 3 4 3 4 S CH₂

1257 1258 1259 1260 3 4 3 4 S CH₂

1261 1262 1263 1264 3 4 3 4 S CH₂

1265 1266 1267 1268 3 4 3 4 S CH₂

1269 1270 1271 1272 3 4 3 4 S CH₂

1273 1274 1275 1276 3 4 3 4 S CH₂

Example 1277 1-[2-(S)-Amino-4-(cyclohexylmethylamino)butanoyl]thiomorpholine dihydrochloride

A. 1-[2-(S)-N-(tert-Butyloxycarbonyl)amino-4-(9-fluorenylmethyloxycarbonylamino)-butanoyl]thiomorpholine

1-[2-(S)-N-(tert-Butyloxycarbonyl)amino-4-(9-fluorenylmethyloxycarbonylamino)-butanoic acid (1.0 g, 2.27 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 20 mL). To this solution at 0° C. were added 1-hydroxybenzotriazole hydrate (461 mg, 3.41 mmol), water-soluble carbodiimide (521 mg, 2.72 mmol), thiomorpholine (281 mg, 2.72 mmol) and triethylamine (340 mg, 3.4 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (100 mL). The solution was washed with 0.3M KHSO₄ (2×25 mL), sat. NaHCO₃ (2×25 mL), water (2×25 mL) and brine (1×25 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 75% ethyl acetate, 25% pet. ether) to give a white solid identified as 1-(2-(S)-N-(tert-butyloxycarbonyl)amino-4-(9-fluorenylmethyloxycarbonylamino)-butanoyl]thiomorpholine (516 mg, 0.98 mmol, 43%).

B. 1-[2-(S)-N-(tert-Butyloxycarbonyl)-4-amino)-butanoyl]thiomorpholine

1-[2-(S)-N-(tert-Butyloxycarbonyl)amino-4-(9-fluorenylmethyloxycarbonylamino)-butanoyl thiomorpholine (500 mg, 0.95 mmol) was dissolved in tetrahydrofuran (20 mL). Diethylamine (5 mL) was added. After 90 min at room temperature the solvent was removed in vacuo and the residue was purified by flash chromatography on silica gel (eluant: 90% chloroform, 7% methanol, 3% triethylamine) to give a pale yellow oil identified as 1-[2-(S)-N-(tert-butyloxycarbonyl)-4-amino)-butanoyl]thiomorpholine (162 mg, 0.54 mmol, 56%).

C. 1-[2-(S)-N-(tert-Butyloxycarbonyl)-amino-4-(cyclohexylmethylamino)butanoyl]thiomorpholine

1-[2-(S)-N-(tert-Butyloxycarbonyl)-4amino)-butanoyl]thiomorpholine (41 mg, 0.135 mmol) was dissolved in dichloroethane (10 mL). To this solution was added cyclohexanecarboxaldehyde (15 mg, 0.135 mmol). After 30 mins sodium triacetoxyborohydride (32 mg, 0.15 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 1% acetic acid,9% methanol, 90% chloroform) to give a colourless oil identified as 1-[2-(S)-N-(tert-butyloxycarbonyl)-amino-4-(cyclohexylmethylamino)butanoyl]thiomorpholine (25 mg, 0.063 mmol, 47%).

D. 1-[2-(S)-Amino-4-(cyclohexylmethylamino)butanoyl]thiomorpholine dihydrochloride

1-[2-(S)-N-(tert-Butyloxycarbonyl)-amino-4-(cyclohexylmethylamino)butanoyl]thiomorpholine (25 mg, 0.063 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 1-[2-(S)-amino-4-(cyclohexylmethylamino)butanoyl]thiomorpholine dihydrochloride (23 mg, 0.063 mmol, 100%).

[M+H]⁺=300.3

Example 1278 1-[2-(S)-Amino-4-((quinolin-2-ylmethyl)amino)butanoyl]thiomorpholine dihydrochloride

A. 1-[2-(S)-N-(tert-Butyloxycarbonyl)-amino-4-((quinolin-2-ylmethyl)amino)butanoyl thiomorpholine

1-[2-(S)-N-(tert-Butyloxycarbonyl)-4-amino)-butanoyl]thiomorpholine (41 mg, 0.135 mmol) was dissolved in 1,2-dichloroethane (10 mL). To this solution was added 2-quinolinecarboxaldehyde (32 mg, 0.15 mmol). After 30 mins sodium triacetoxyborohydride (36 mg, 0.17 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 1% acetic acid,9% methanol, 90% chloroform) to give a colourless oil identified as 1-[2-(S)-N-(tert-butyloxycarbonyl)-amino-4-((quinolin-2-ylmethyl)amino)butanoyl thiomorpholine (32 mg, 0.072 mmol, 53%).

B. 1-[2-(S)-Amino-4-((quinolin-2-ylmethyl)amino)butanoyl]thiomorpholine dihydrochloride

1-[2-(S)-N-(tert-Butyloxycarbonyl)-amino-4-((quinolin-2-ylmethyl)amino)butanoyl]thiomorpholine (12 mg, 0.027 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 4 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 1-[2-(S)-amino-4-((quinolin-2-ylmethyl)amino)butanoyl]thiomorpholine dihydrochloride (11.3 mg, 0.027 mmol, 100%).

[M+H]⁺=345.3

Example 1279 1-[2-(S)-Amino-4-(cyclohexylmethylamino)butanoyl]piperidine dihydrochloride

A. 1-[2-(S)-N-(tert-Butyloxycarbonyl)amino-4-(9-fluorenylmethyloxycarbonylamino)-butanoyl]piperidine

1-[2-(S)-N-(tert-Butyloxycarbonyl)amino-4-(9-fluorenylmethyloxycarbonylamino)-butanoic acid (947 mg, 2.154 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 20 mL). To this solution at 0° C. were added 1-hydroxybenzotriazole hydrate (436 mg, 3.2 mmol), water-soluble carbodiimide (495g, 2.58 mmol), piperidine (220 g, 2.58 mmol) and triethylamine (320 mg, 3.2 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (100 mL). The solution was washed with 0.3M KHSO₄ (2×25 mL), sat. NaHCO₃ (2×25 mL), water (2×25 mL) and brine (1×25 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 75% ethyl acetate, 25% pet. ether) to give a white solid identified as 1-[2-(S)-N-(tert-butyloxycarbonyl)amino-4-(9-fluorenylmethyloxycarbonylamino)-butanoyl]piperidine (556 mg, 1.1 mmol, 51%).

B. 1-[2-(S)-N-(tert-Butyloxycarbonyl)-4-amino)-butanoyl]piperidine

1-[2-(S)-N-(tert-Butyloxycarbonyl)amino-4-(9-fluorenylmethyloxycarbonylamino)-butanoyl]piperidine (540 g, 1.1 mmol) was dissolved in tetrahydrofuran (20 mL). Diethylamine (5 mL) was added. After 90 min at room temperature the solvent was removed in vacuo and the residue was purified by flash chromatography on silica gel (eluant: 90% chloroform, 7% methanol, 3% triethylamine) to give a pale yellow oil identified as 1-[2-(S)-N-(tert-butyloxycarbonyl)-4-amino)-butanoyl]piperidine (171 mg, 0.6 mmol, 57%).

C. 1-[2-(S)-N-(tert-Butyloxycarbonyl)-amino-4-(cyclohexylmethylamino)butanoyl]piperidine

1-[2-(S)-N-(tert-Butyloxycarbonyl)-4-amino)-butanoyl]piperidine (43 mg, 0.15 mmol) was dissolved in 1,2-dichloroethane (20 mL). To this solution was added cyclohexanecarboxaldehyde (17 mg, 0.15 mmol). After 30 mins sodium triacetoxyborohydride (36 mg, 0.17 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 1% acetic acid,9% methanol, 90% chloroform) to give a colourless oil identified as 1-[2-(S)-N-(tert-butyloxycarbonyl)-amino-4-(cyclohexylmethylamino)butanoyl]piperidine (38 mg, 0.1 mmol, 66%).

D. 1-[2-(S)-Amino-4-(cyclohexylmethylamino)butanoyl]piperidine dihydrochloride

1-[2-(S)-N-(tert-Butyloxycarbonyl)-amino-4-(cyclohexylmethylamino)butanoyl]piperidine (38 mg, 0.1 mmol) was dissolved in 4M HCl/dioxan (2 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 1-[2-(S)-amino-4-(cyclohexylmethylamino)butanoyl]piperidine dihydrochloride (33 mg, 0.093 mmol, 93%).

[M+H]⁺=282.3

Example 1280 1-[2-(S)-Amino-4-((quinolin-2-ylmethyl)amino)butanoyl]piperidine dihydrochloride

A. 1-[2-(S)-N-(tert-Butyloxycarbonyl)-amino-4-((quinolin-2-ylmethyl)amino)butanoyl]piperidine

1-[2-(S)-N-(tert-Butyloxycarbonyl)-4-amino)-butanoyl]piperidine (24 mg, 0.15 mmol) was dissolved in 1,2-dichloroethane (25 mL). To this solution was added 2-quinolinecarboxaldehyde (24 mg, 0.15 mmol). After 30 mins sodium triacetoxyborohydride (36 mg, 0.17 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 1% acetic acid,9% methanol, 90% chloroform) to give a colourless oil identified as 1-[2-(S)-N-(tert-butyloxycarbonyl)-amino-4-((quinolin-2-ylmethyl)amino)butanoyl]piperidine (35 mg, 0.082 mmol, 55%).

B. 1-[2-(S)-Amino-4-((quinolin-2-ylmethyl)amino)butanoyl]piperidine dihydrochloride

1-[2-(S)-N-(tert-Butyloxycarbonyl)-amino-4-((quinolin-2-ylmethyl)amino)butanoyl]piperidine (35 mg, 0.082 mmol) was dissolved in 4M HCl/dioxan (2 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 1-[2-(S)-amino-4-((quinolin-2-ylmethyl)amino)butanoyl]piperidine dihydrochloride (26 mg, 0.065 mmol, 79%).

[M+H]⁺=327.3

Example 1281 3-Fluoro-1-[2-(S)-amino-4-(cyclohexenylmethylamino)butanoyl]pyrrolidine dihydrochloride

A. 1-(tert-Butyloxycarbonyl)-3-fluoropyrrolidine

N-(tert-Butyloxycarbonyl)-3-hydroxypyrrolidine (21.0 g, 10.7 mmol) was dissolved in CH₂Cl₂ (30 ml). (Diethylamino)sulphur trifluoride (1.72 g, 10.7 mmol) was added to this solution at −78° C. The mixture was stirred for 18 hours at −78° C. to room temperature then the reaction mixture was carefully poured into sat. NaHCO₃ (100 ml) and stirred for 15 min and extracted with CH₂Cl₂. The organic extract was washed with water and brine, dried (Na₂SO₄) and evaporated in vacuo to give an orange oil. The residue was purified by flash chromatography (eluant: 28% ethyl acetate, 72% pet. ether 60-80) to give a colourless oil identified as 1-(tert-butyloxycarbonyl)-3-fluoropyrrolidine (1.14 g, 5.34 mmol, 50%).

B. 3-Fluoropyrrolidine hydrochloride

1-(tert-Butyloxycarbonyl)-3-fluoropyrrolidine (1.14 g, 5.34 mmol) was dissolved in 4M HCl/dioxan (30 ml). The mixture was stirred for 1 hour at room temperature then the solvent was removed in vacuo to give an off-white solid identified as 3-fluoropyrrolidine hydrochloride (640 mg, 5.2 mmol, 95%).

C. 3-Fluoro-1-[2-(S)-N-(tert-butyloxycarbonyl)amino-4-(9-fluorenylmethyloxycarbonylamino)-butanoyl]pyrrolidine

1-[2-(S)-N-(tert-Butyloxycarbonyl)amino-4-(9-fluorenylmethyloxycarbonylamino)-butanoic acid (950 mg, 2.15 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 20 mL). To this solution at 0° C. were added 1-hydroxybenzotriazole hydrate (395 mg, 2.6 mmol), water-soluble carbodiimide (572 mg, 3.0 mmol), 3-fluoropyrrolidine hydrochloride (270 g, 2.15 mmol) and triethylamine (320 mg, 3.2 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (100 mL). The solution was washed with 0.3M KHSO₄ (2×25 mL), sat. NaHCO₃ (2×25 mL), water (2×25 mL) and brine (1×25 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 75% ethyl acetate, 25% pet. ether) to give a white solid identified as 3-fluoro1-[2-(S)-N-(tert-butyloxycarbonyl)amino-4-(9-fluorenylmethyloxycarbonylamino)-butanoyl]pyrrolidine (808 mg, 1.58 mmol, 73%).

D. 3-Fluoro-1-[2-(S)-N-(tert-butyloxycarbonyl)-4-amino)-butanoyl]pyrrolidine

3-Fluoro-1-[2-(S)-N-(tert-butyloxycarbonyl)amino-4-(9-fluorenylmethyloxycarbonylamino)-butanoyl]pyrrolidine (800 mg, 1.58 mmol) was dissolved in tetrahydrofuran (20 mL). Diethylamine (5 mL) was added. After 90 min at room temperature the solvent was removed in vacuo and the residue was purified by flash chromatography on silica gel (eluant: 90% chloroform, 7% methanol, 3% triethylamine) to give a pale yellow oil identified as 3-fluoro-1-[2-(S)-N-(tert-butyloxycarbonyl)-4-amino)-butanoyl]pyrrolidine (316 mg, 1.04 mmol, 66%).

E. 3-Fluoro-1-[2-(S)-N-(tert-butyloxycarbonyl)-amino-4-(cyclohexenylmethylamino)butanoyl]pyrrolidine

3-Fluoro-1-(2-(S)-N-(tert-butyloxycarbonyl)-4-amino)-butanoyl]pyrrolidine (150 mg, 0.52 mmol) was dissolved in methanol (20 mL). To this solution was added 3-cyclohexenecarboxaldehyde (63 mg, 0.57 mmol). After 30 mins sodium triacetoxyborohydride (220 mg, 1.04 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 1% acetic acid,9% methanol, 90% chloroform) to give a colourless oil identified as 3-fluoro-1-[2-(S)-N-(tert-butyloxycarbonyl)-amino-4-(cyclohexenylmethylamino)butanoyl]pyrrolidine (176 mg, 0.46 mmol, 77%).

F. 3-Fluoro-1-[2-(S)-amino-4-(cyclohexenylmethylamino)butanoyl]pyrrolidine dihydrochloride

3-Fluoro-1-[2-(S)-N-(tert-butyloxycarbonyl)-amino-4-(cyclohexenylmethylamino)butanoyl]pyrrolidine (176 mg, 0.46 mmol) was dissolved in 4M HCl/dioxan (2 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 3-fluoro-1-[2-(S)-amino-4-(cyclohexenylmethylamino)butanoyl]pyrrolidine dihydrochloride (140 mg, 0.39 mmol, 963%).

[M+H]⁺=284.3

Example 1282 1-[2-(S)-Amino-4-(N-methyl-N-(2-methylbenzyl)amino)butanoyl]piperidine dihydrochloride

A. N-(tert-Butyloxycarbonyl)-L-homoserine lactone

L-Homoserine lactone 1.76 g, 12.8 mmol) was dissolved in DMF (30 mL). This solution was cooled to 0° C., triethylamine (1.41, 14.1 mmol) di-tert-butyl dicarbonate(3.35 g, 15.35 mmol) was added. After 18 hours at room temperature the solvent was evaporated in vacuo, the residue was taken up in dichloromethane (200 mL). This solution was washed with 1M KHSO₄ (2×50 mL) and brine (1×50 mL), dried (Na₂SO₄) and evaporated in vacuo to give a white solid, recrystallised from EtOAc/pet.ether to give a white solid identified as N-(tert-butyloxycarbonyl)-L-homoserine lactone (2.25 mg, 11.2 mmol, 87%).

B. 1-[2-(S)-(N-(tert-Butyloxycarbonyl)amino)-4-hydroxybutanoyl]piperidine

N-(tert-Butyloxycarbonyl)-L-homoserine lactone (100 mg, 0.5 mmol) was dissolved in tetrahydrofuran (30 mL). Piperidine (42 mg, 0.5 mmol) was added. After 72 hours at room temperature the reaction mixture was diluted with ethyl acetate (150 mL). This solution was washed with water (1×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil identified as 1-[2-(S)-(N-(tert-butyloxycarbonyl)amino)-4-hydroxybutanoyl]piperidine (142 mg, 0.5 mmol, 100%).

C. 1-[2-(S)-(N-(tert-Butyloxycarbonyl)amino)-4-oxobutanoyl]piperidine

1-[2-(S)-(N-(tert-Butyloxycarbonyl)amino)-4-hydroxybutanoyl]piperidine (142 mg, 0.5 mmol) was dissolved in dichloromethane (50 mL). Dess-Martin periodinane (232 mg, 0.5 mmol) was added. After 1 hour at room temperature the reaction mixture was diluted with ethyl acetate (150 mL). This solution was washed with water (1×20 ml) and brine (1×20 ml), dried (Na₂SO₄) and evaporated in vacuo to give a colourless oil. Purified by flash chromatography on silica gel (eluant: 50% ethyl acetate, 50% pet. ether 60-80) to give a colourless oil identified as 1-[2-(S)-(N-(tert-butyloxycarbonyl)amino)-4-oxobutanoyl]piperidine (40 mg, 0.14 mmol, 27%).

D. 1-[2-(S)-(N-(tert-butyloxycarbonyl)amino-4-(N-methyl-N-(2-methylbenzyl)amino)butanoyl]piperidine

1-[2-(S)-(N-(tert-Butyloxycarbonyl)amino)-4-oxobutanoyl]piperidine (40 mg, 14 mmol) was dissolved in methanol (20 mL). To this solution was added N-methyl-2-methylbenzylamine (19 mg, 0.14 mmol). After 2 hours sodium triacetoxyborohydride (64 mg, 0.3 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel to give a colourless oil identified as 1-[2-(S)-(N-(tert-butyloxycarbonyl)amino-4-(N-methyl-N-(2-methylbenzyl)amino)butanoyl]piperidine (36 mg, 0.09 mmol, 64%).

E. 1-[2-(S)-Amino-4-(N-methyl-N-(2-methylbenzyl)amino)butanoyl]piperidine dihydrochloride

1-[2-(S)-(N-(tert-Butyloxycarbonyl)amino-4-(N-methyl-N-(2-methylbenzyl)amino)butanoyl]piperidine (36 mg, 0.09 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a pale brown solid identified as 1-[2-(S)-amino-4-(N-methyl-N-(2-methylbenzyl)amino)butanoyl]piperidine dihydrochloride (43 mg, 0.09 mmol, 100%)

Example 1283 1-[N-(2″-(Cyclohexylmethylaminocthyl)glycinyl)]thiomorpholine dihydrochloride

A. 1-[N-2′-(tert-Butyloxycarbonyl)-N-(2″-(9-fluorenylmethyloxycarbonyl aminoethyl)-glycinyl]thiomorpholine

N-2′-(tert-Butyloxycarbonyl)-N-(2″-(9-fluorenylmethyloxycarbonyl aminoethyl)-glycine (2.5 g, 5.7 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 100 mL). To this solution at 0° C. were added 1-hydroxybenzotriazole hydrate (833 mg, 6.3 mmol), water-soluble carbodilmide (974 mg, 6.3 mmol), thiomorpholine (617 mg, 6.0 mmol) and N-methylmorpholine (800 mg, 8 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (100 mL). The solution was washed with 0.3M KHSO₄ (2×25 mL), sat. NaHCO₃ (2×25 mL), water (2×25 mL) and brine (1×25 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 75% ethyl acetate, 25% pet. ether) to give a white solid identified as 1-[N-2′-(tert-butyloxycarbonyl)-N-(2″-(9-fluorenylmethyloxycarbonyl aminoethyl)-glycinyl]thiomorpholine (2.7 g, 5.1 mmol, 90%).

B. 1-[N-2′-(tert-Butyloxycarbonyl)-(2″-aminoethyl)-glycinyl]thiomorpholine

1-[N-2′-(tert-Butyloxycarbonyl)-N-(2″-(9-fluorenylmethyloxycarbonyl aminoethyl)-glycinyl]thiomorpholine (2.7 g, 5.1 mmol) was dissolved in tetrahydrofuran (20 mL). Diethylamine (5 mL) was added. After 90 min at room temperature the solvent was removed in vacuo and the residue was purified by flash chromatography on silica gel (eluant: 90% chloroform, 7% methanol, 3% triethylamine) to give a pale yellow oil identified as 1-[N-2′-(tert-butyloxycarbonyl)-(2″-aminoethyl)-glycinyl]thiomorpholine (1.44 g, 4.7 mmol, 92%).

C. 1-[2′-N-(tert-Butyloxycarbonyl N-(2″-(cyclohexylmethylaminoethyl)-glycinyl]thiomorpholine

1-[N-2′-(tert-Butyloxycarbonyl)-(2″-aminoethyl)-glycinyl]thiomorpholine (100 mg, 0.3 mmol) was dissolved in methanol (25 mL). To this solution was added cyclohexanecarboxaldehyde (34 mg, 0.3 mmol). After 30 mins sodium triacetoxyborohydride (126 mg, 0.6 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 1% acetic acid,9% methanol, 90% chloroform) to give a colourless oil identified as 1-[2′-N-(tert-Butyloxycarbonyl N-(2″-(cyclohexylmethylaminoethyl)-glycinyl]thiomorpholine (33 mg, 0.08 mmol, 27%).

D. 1-[N-(2″-(Cyclohexylmethylaminoethyl)glycinyl)]thiomorpholine dihydrochloride

1-[2′-N-(tert-Butyloxycarbonyl-N-(2″-(cyclohexylmethylaminoethyl)-glycinyl]thiomorpholine (33 mg, 0.081 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 1-[N-(2″-(cyclohexylmethylaminoethyl)glycinyl)]thiomorpholine dihydrochloride (31 mg, 0.08 mmol, 100%).

[M+H]⁺=300.3

Example 1284 1-[N-(2″-((Quinolin-2-ylmethyl)aminoethyl)glycinyl)]pyrrolidine dihydrochloride

A. 1-[N-2′-(tert-Butyloxycarbonyl)-N-(2″-(9-fluorenylmethyloxycarbonyl aminoethyl)-glycinyl]piperidine

N-2′-(tert-Butyloxycarbonyl)-N-(2″-(9-fluorenylmethyloxycarbonyl aminoethyl)-glycine (2.5 g, 5.7 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 100 mL). To this solution at 0° C. were added 1-hydroxybenzotriazole hydrate (1.5 g, 11.1 mmol), water-soluble carbodiimide (1.3 g, 6.8 mmol), piperidine (484 mg, 5.69 mmol) and N-methylmorpholine (800 mg, 8 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (100 mL). The solution was washed with 0.3M KHSO₄ (2×25 mL), sat. NaHCO₃ (2×25 mL), water (2×25 mL) and brine (1×25 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 75% ethyl acetate, 25% pet. ether) to give a white solid identified as 1-[N-2′-(tert-butyloxycarbonyl)-N-(2″-(9-fluorenylmethyloxycarbonyl aminoethyl)-glycinyl]piperidine (2.8 g, 5.5 mmol, 96%).

B. 1-[N-2′-(tert-Butyloxycarbonyl)-(2″-aminoethyl)-glycinyl]piperidine

1-[N-2′-(tert-Butyloxycarbonyl)-N-(2″-(9-fluorenylmethyloxycarbonyl aminoethyl)-glycinyl]piperidine (2.8 g, 5.5 mmol) was dissolved in tetrahydrofuran (20 mL). Diethylamine (5 mL) was added. After 90 min at room temperature the solvent was removed in vacuo and the residue was purified by flash chromatography on silica gel (eluant: 90% chloroform, 7% methanol, 3% triethylamine) to give a pale yellow oil identified as 1-[N-2′-(tert-butyloxycarbonyl)-(2″-aminoethyl)-glycinyl]piperidine (1.4 g, 4.9 mmol, 89%).

C. 1-[2′-N-(tert-Butyloxycarbonyl N-(2″-((quinolin-2-ylmethyl)aminoethyl)-glycinyl]piperidine

1-[N-2′-(tert-Butyloxycarbonyl)-(2″-aminoethyl)-glycinyl]piperidine was dissolved in methanol (25 mL). To this solution was added 2-quinolinecarboxaldehyde. After 30 mins sodium triacetoxyborohydride was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 1% acetic acid,9% methanol, 90% chloroform) to give a colourless oil identified as 1-[2′-N-(tert-butyloxycarbonyl N-(2″-((quinolin-2-ylmethyl)aminoethyl)-glycinyl]piperidine.

D. 1-[N-(2″-((Quinolin-2-ylmethyl)aminoethyl)glycinyl)]piperidine dihydrochloride

1-2′-N-(tert-Butyloxycarbonyl-N-(2″-((quinolin-2-ylmethyl)aminoethyl)-glycinyl]piperidine was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 1-[N-(2″-((quinolin-2-ylmethyl)aminoethyl)glycinyl)]piperidine dihydrochloride.

Example 1285 1-[N,N-(2″,2″-((Dicinnamyl)aminoethyl)glycinyl)]thiomorpholine dihydrochloride

A. 1-[2′-N-(tert-Butyloxycarbonyl N,N-(2″,2″-((dicinnamyl)aminoethyl)-glycinyl]thiomorpholine

(2S)-1-(N^(α)-(tert-Butyloxycarbonyl)-L-lysinyl)-pyrrolidine-2-carbonitrile (250 mg, 0.83 mmol) was dissolved in dichloroethane (25 mL). To this solution was added trans-cinnamaldehyde (108 mg, 0.83 mmol). After 30 mins sodium triacetoxyborohydride (350 mg, 1.6 mmol) was added. After 18 h at room temperature the solvent was removed in vacuo and the residue was taken up in chloroform (70 mL). This solution was washed with water (2×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography on silica gel (eluant: 2% methanol, 98% chloroform) to give a colourless oil identified as 1-[2′-N-(tert-butyloxycarbonyl N,N-(2″,2″-((dicinnamyl)aminoethyl)-glycinyl]thiomorpholine. Further elution with 9% methanol, 90% chloroform and 1% acetic acid gave a colourless oil identified as 1-[2′-N-(tert-butyloxycarbonyl N,-(2″-((cinnamyl)aminoethyl)glycinyl]thiomorpholine (180 mg, 0.43 mmol, 52%)

B. 1-[N,N-(2″,2″-((Dicinnamyl)aminoethyl)glycinyl)]thiomorpholine dihydrochloride

1-[2′-N-(tert-Butyloxycarbonyl N,N-(2″,2″-((dicinnamyl)aminoethyl)-glycinyl]thiomorpholine was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 1-[N,N-(2″,2″-((dicinnamyl)aminoethyl)glycinyl)]thiomorpholine dihydrochloride.

Example 1286 1-[N-(2″-((Cinnamyl)aminoethyl)glycinyl)]thiomorpholine dihydrochloride

A. 1-[N-(2″-((Cinnamyl)aminoethyl)glycinyl)]thiomorpholine dihydrochloride

1-[2′-N-(tert-Butyloxycarbonyl N-(2″-((cinnamyl)aminoethyl)-glycinyl]thiomorpholine (180 mg, 0.43 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 1-[N-(2″-((cinnamyl)aminoethyl)glycinyl)]thiomorpholine dihydrochloride (168 mg, 0.43 mmol, 100%).

[M+H]⁺=320.3

Example 1287 3,3-Difluoro-1-[N-2″-(3′-trifluoromethylanilino)pyridyl-3-carbonyl aminoethyl)glycinyl)]pyrrolidine dihydrochloride

A. 3,3-Difluoro-1-[N-2′-(tert-butyloxycarbonyl)-N-(2″-(9-fluorenylmethyloxycarbonyl aminoethyl)-glycinyl]pyrrolidine

N-2′-(tert-Butyloxycarbonyl)-N-(2″-(9-fluorenylmethyloxycarbonyl aminoethyl)-glycine (1.0 g, 2.27 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 100 mL). To this solution at 0° C. were added 1-hydroxybenzotriazole hydrate (620 mg, 4.6 mmol), water-soluble carbodiimide (560 mg, 2.8 mmol), 3,3-difluoropyrrolidine hydrochloride (360 mg, 2.5 mmol) and N-methylmorpholine (800 mg, 8 mmol). After 18h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (100 mL). The solution was washed with 0.3M KHSO₄ (2×25 mL), sat. NaHCO₃ (2×25 mL), water (2×25 mL) and brine (1×25 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 60% ethyl acetate, 40% pet. ether) to give a white solid identified as 3,3-difluoro-1-[N-2′-(tert-butyloxycarbonyl)-N-(2″-(9-fluorenylmethyloxycarbonyl aminoethyl)-glycinyl]pyrrolidine (934 g, 1.7 mmol, 77%).

B.3,3-Difluoro-1-[N-2′-(tert-butyloxycarbonyl)aminoethyl)-glycinyl]pyrrolidine

3,3-Difluoro-1-[N-2′-(tert-butyloxycarbonyl)-N-(2″-(9-fluorenylmethyloxycarbonyl aminoethyl)-glycinyl]pyrrolidine (890 g, 1.68 mmol) was dissolved in tetrahydrofuran (20 mL). Diethylamine (5 mL) was added. After 90 min at room temperature the solvent was removed in vacuo and the residue was purified by flash chromatography on silica gel (eluant: 90% chloroform, 7% methanol, 3% triethylamine) to give a pale yellow oil identified as 3,3-difluoro-1-[N-2′-(tert-butyloxycarbonyl)aminoethyl)-glycinyl]pyrrolidine (470 mg, 1.5 mmol, 91%).

C. 3,3-Difluoro-1-N-2′-(tert-butyloxycarbonyl)-N-2″-(3′-trifluoromethylanilino)pyridyl-3-carbonyl aminoethyl)glycinyl)]pyrrolidine

3,3-Difluoro-1-[N-2′-(tert-butyloxycarbonyl)aminoethyl)-glycinyl]pyrrolidine (50 mg, 0.16 mmol) was dissolved in CH₂Cl₂/DMF (9:1, 20 mL). To this solution at 0° C. was added 1-hydroxybenzotriazole hydrate (46 mg, 0.34 mmol), water-soluble carbodiimide (40 mg, 0.2 mmol), niflumic acid (49 mg, 0.17 mmol) and N-methylmorpholine (40 mg, 0.4 mmol). After 18 h at 0° C. to room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (70 mL). The solution was washed with 0.3M KHSO₄ (1×20 mL), sat. NaHCO₃ (1×20 mL), water (1×20 mL) and brine (1×20 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was purified by flash chromatography on silica gel (eluant: 75% ethyl acetate, 25% pet. ether) to give a yellow oil identified as 3,3-difluoro-1-[N-2′-(tert-butyloxycarbonyl)-N-2″-(3′-trifluoromethylanilino)pyridyl-3-carbonyl aminoethyl)glycinyl)]pyrrolidine (63 mg, 0.11 mmol, 67%).

D. 3,3-Difluoro-1-[N-2″-(3′-trifluoromethylanilino)pyridyl-3-carbonyl aminoethyl) glycinyl)]pyrrolidine dihydrochloride

3,3-Difluoro-1-[N-2′-(tert-butyloxycarbonyl)-N-2″-(3′-trifluoromethylanilino)pyridyl-3-carbonyl aminoethyl)glycinyl)]pyrrolidine (55 mg, 0.10 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a pale brown solid identified as 3,3-difluoro-1-[N-2″-(3′-trifluoromethylanilino)pyridyl-3-carbonyl aminoethyl)glycinyl)]pyrrolidine dihydrochloride (52 mg, 0.10 mmol, 100%).

[M+H]⁺=472.3

Example 1288 3,3-Difluoro-[N-2″-(6-Chloro-4-(4′-fluoroanilino)-1,3,5-triazinyl)aminoethyl) glycinyl)]thiomorpholine dihydrochloride

A. 4,6-Dichloro-2-(4′-fluoroanilino)-1,3,5-triazine

Cyanuric chloride (1.844 g, 10 mmol) was dissolved in acetonitrile (20 mL). The solution was cooled to −20° C. A solution of 4-fluoroaniline (1.1 g, 10 mmol) and triethylamine (1.0 g, 10 mmol) was slowly added. After 1 hour at −20° C. the solvent was removed in vacuo and the residue was taken up in ethyl acetate (150 mL). The solution was washed with water (1×50 mL) and brine (1×50 mL), dried (Na₂SO₄) and evaporated in vacuo. The residue was recrystallised from ethyl acetate/hexane to give an off white solid identified as 4,6-dichloro-2-(4′-fluoroanilino)-1,3,5-triazine 1.7 g, 6.0 mmol, 60%).

B. 1-[N-2′-(tert-butyloxycarbonyl)-N-2″-(6-Chloro-4-(4′-fluoroanilino)-1,3,5-triazinyl aminoethyl)glycinyl)]thiomorpholine

1-[N-2′-(tert-butyloxycarbonyl)aminoethyl)-glycinyl]thiomorpholine (100 mg, 0.3 mmol) was dissolved in dichloromethane (30 mL). To this solution was added 4,6-dichloro-2-(4′-fluoroanilino)-1,3,5-triazine (90 mg, 0.3 mmol) and triethylamine (50 mg, 0.5 mmol). After 2 hours at room temperature the solvent was removed in vacuo and the residue was taken up in ethyl acetate (150 mL). This solution was washed with water (2×30 mL) and brine (1×30 mL), dried (Na₂SO₄) and evaporated in vacuo to give a yellow oil. The residue was purified by flash chromatography (eluant: 60% ethyl acetate, 40% pet. ether) to give a white solid identified as 1-[N-2′-(tert-butyloxycarbonyl)-N-2″-(6-chloro-4-(4′-fluoroanilino)1,3,5-triazinyl aminoethyl)glycinyl)]thiomorpholine (20 mg, 0.032 mmol, 11%).

C. 1-[N-2″-(6-Chloro-4-(4′-fluoroanilino)-1,3,5-triazinyl)aminoethyl)glycinyl)]thiomorpholine dihydrochloride

1-[N-2′-(tert-butyloxycarbonyl)-N-2″-(6-chloro-4-(4′-fluoroanilino)-1,3,5-triazinyl aminoethyl)glycinyl)]thiomorpholine (18.8 mg, 0.03 mmol) was dissolved in 4M HCl/dioxan (20 mL). After 1 h at room temperature the solvent was removed in vacuo. The residue was lyophilised from water to give a white solid identified as 1-[N-2″-(6-Chloro-4-(4′-fluoroanilino)-1,3,5-triazinyl)aminoethyl)glycinyl)]thiomorpholine dihydrochloride (18 mg, 0.03 mmol, 100%).

[M+H]⁺=526.4 TABLE 9

Ex No X a R 1289 1290 1291 1292 1293 1294 S CF₂CHF S CH₂O 1  2

1295 1296 1297 1298 1299 1300 S CF₂CHF S CH₂O 1  2

1311 1312 1313 1314 1315 1316 S CF₂CHF S CH₂O 1  2

1317 1318 1319 1320 S CF₂CHF O 1  2

1321 1322 1323 1324 1325 1326 S CF₂CHF S CH₂O 1  2

1327 1328 1329 1330 1331 1332 S CF₂CHF S CH₂O 1  2

1333 1334 1335 1336 1337 1338 S CF₂CHF S CH₂O 1  2

1339 1340 1341 1342 1343 1344 S CF₂CHF S CH₂O 1  2

1345 1346 1347 1348 1349 1350 S CF₂CHF S CH₂O 1  2

1351 1352 1353 1354 1355 1356 S CF₂CHF S CH₂O 1  2

1357 1358 1359 1360 1361 1362 S CF₂CHF S CH₂O 1  2

1363 1364 1365 1366 1367 1368 S CF₂CHF S CH₂O 1  2

1369 1370 1371 1372 1373 1374 S CF₂CHF S CH₂O 1  2

1375 1376 1377 1378 1379 1380 S CF₂CHF S CH₂O 1  2

1381 1382 1383 1384 1385 1386 S CF₂CHF S CH₂O 1  2

1387 1388 1389 1390 1391 1392 S CF₂CHF S CH₂O 1  2

1393 1394 1395 1396 1397 1398 S CF₂CHF S CH₂O 1  2

1399 1400 1401 1402 1403 1404 S CF₂CHF S CH₂O 1  2

1405 1406 1407 1408 1409 1410 S CF₂CHF S CH₂O 1  2

1411 1412 1413 1414 1415 1416 S CF₂CHF S CH₂O 1  2

1417 1418 1419 1420 1421 1422 S CF₂CHF S CH₂O 1  2

1423 1424 1425 1426 1427 1428 S CF₂CHF S CH₂O 1  2

1429 1430 1431 1432 1433 1434 S CF₂CHF S CH₂O 1  2

TABLE 10

Ex No X a R 1614 1615 1616 1617 S CF₂S CH₂ 1 2

1618 1619 1620 1621 S CF₂S CH₂ 1 2

1622 1623 1624 1625 S CF₂S CH₂ 1 2

1626 1627 1628 1629 S CF₂S CH₂ 1 2

1630 1631 1632 1633 S CF₂S CH₂ 1 2

1634 1635 1636 1637 S CF₂S CH₂ 1 2

1638 1639 1640 S CF₂S CH₂ 1 2

1642 1643 1644 1645 S CF₂S CH₂ 1 2

1646 1647 1648 1649 S CF₂S CH₂ 1 2

1650 1651 1652 1653 S CF₂S CH₂ 1 2

1654 1655 1656 1657 S CF₂S CH₂ 1 2

1658 1659 1660 1661 S CF₂S CH₂ 1 2

1662 1663 1664 1665 S CF₂S CH₂ 1 2

1666 1667 1668 1669 S CF₂S CH₂ 1 2

1670 1671 1672 1673 S CF₂S CH₂ 1 2

1674 1675 1676 1677 S CF₂S CH₂ 1 2

1678 1679 1680 1681 S CF₂S CH₂ 1 2

1682 1683 1684 1685 S CF₂S CH₂ 1 2

1686 1687 1688 1689 S CF₂S CH₂ 1 2

1690 1691 1692 1693 S CF₂S CH₂ 1 2

1694 1695 1696 1697 S CF₂S CH₂ 1 2

1698 1699 1700 1701 S CF₂S CH₂ 1 2

1702 1703 1704 1705 S CF₂S CH₂ 1 2 

1. A compound according to general formula 1, or a pharmaceutically acceptable salt thereof,

wherein: either G¹ is —CH₂—X²—(CH₂)_(a)—G³ and G² is H, or G2 is —CH₂—(CH₂)_(a)—G³ and G¹ is H; G³ is selected from a group according to general formula 2, a group according to general formula 3, and a group according to general formula 4;

a is 0, 1 or 2; b is 1 or 2; X¹ is selected from CH₂, S, CF₂, CHF, CH(CH₃), C(CH₃)₂, CH(CN) and O; X² is selected from CH₂, O and S, provided that if a is 1 then X² is CH₂; X³, X⁴ and X⁵ are selected from N and CH, provided that at least two of X³, X⁴ and X⁵ are N; X⁶ is selected from O and NH; X⁷ is selected from CH₂, O, S and NH; R¹ is selected from H and CN; R² is selected from H and alkyl; R³ is selected from H, Cl, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂; R⁴, R⁵, R⁶, R⁷ and R⁸ are independently selected from H, Br, Cl, F, CF₃, alkyl, acyl, OH, O-alkyl, NH₂, NH-alkyl, N(alkyl)₂, NO₂, NH-acyl, CO₂H, CO₂-alkyl, CONH₂, CONH-alkyl, CON(alkyl)₂ and CN; R⁹ is selected from H and alkyl; R¹⁰, R¹¹, R¹², R¹³ and R¹⁴ are independently selected from H, Br, Cl, F, CF₃, alkyl, acyl, OH, O-alkyl, NH₂, NH-alkyl, N(alkyl)₂, NO₂, NH-acyl, CO₂H, CO₂-alkyl, CONH₂, CONH-alkyl, CON(alkyl)₂ and CN; R¹⁵ and R¹⁶ are independently selected from H, alkyl, alkenyl, polyfluoroalkyl, aralkyl, aryl and —CH₂—L—R¹⁷, or R¹⁵ and R¹⁶ together form a group according to general formula 5, general formula 6 or general formula 7;

R¹⁷ is selected from H, alkyl and aryl; R¹⁸ is selected from H, alkyl, aryl, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂; R¹⁹ is selected from H, alkyl, aryl, F, Cl, Br, CF₃, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂; L is selected from a covalent bond, CH═CH, C≡C and —C₆H₄—; d and e are selected from 0, 1, 2 and 3 such that d+e is 3,4 or 5; and f is selected from 1, 2 and 3; provided that when R¹⁵ and R¹⁶ are both H and b is 1 then X¹ is not S or CH₂.
 2. A compound according to general formula 8, or a pharmaceutically acceptable salt thereof,

wherein: a is 0, 1 or 2; b is 1 or 2; X¹ is selected from CH₂, S, CF₂, CHF, CH(CH₃)₂, C(CH₃)₂, CH(CN) and O; X² is selected from CH₂, O and S, provided that if a is 1 then X² is CH₂; X³, X⁴ and X⁵ are selected from N and CH, provided that at least two of X³, X⁴ and X⁵ are N; X⁶ is selected from O and NH; R¹ is selected from H and CN; R² is selected from H and alkyl; R³ is selected from H, Cl, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂; R⁴, R⁵, R⁶, R⁷ and R⁸ are independently selected from H, Br, Cl, F, CF₃, alkyl, acyl, OH, O-alkyl, NH₂, NH-alkyl, N(alkyl)₂, NO₂, NH-acyl, CO₂H, CO₂-alkyl, CONH₂, CONH-alkyl, CON(alkyl)₂ and CN.
 3. A compound according to claim 2 wherein R¹ is H.
 4. A compound according to claim 2 wherein R¹ is CN.
 5. A compound according to claim 2 wherein X¹ is CH₂.
 6. A compound according to claim 2 wherein X¹ is S.
 7. A compound according to claim 2 wherein b is
 1. 8. A compound according to claim 2 wherein b is
 2. 9. A compound according to claim 2 wherein a is
 1. 10. A compound according to claim 2 wherein a is 2 and X² is CH₂.
 11. A compound according to claim 2 wherein X³, X⁴ and X⁵ are all N.
 12. A compound according to general formula 9, or a pharmaceutically acceptable salt thereof,

wherein: a is 1 or 2; b is 1 or 2; X¹ is selected from CH₂, S, CF₂, CHF, CH(CH₃)₂, C(CH₃)₂, CH(CN) and O; X³, X⁴ and X⁵ are selected from N and CH, provided that at least two of X³, X⁴ and X⁵ are N; X⁶ is selected from O and NH; R¹ is selected from H and CN; R² is selected from H and alkyl; R³ is selected from H, Cl, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂; R⁴, R⁵, R⁶, A⁷ and R⁸ are independently selected from H, Br, Cl, F, CF₃, alkyl, acyl, OH, O-alkyl, NH₂, NH-alkyl, N(alkyl)₂, NO₂, NH-acyl, CO₂H, CO₂-alkyl, CONH₂, CONH-alkyl, CON(alkyl)₂ and CN.
 13. A compound according to claim 12 wherein R¹ is H.
 14. A compound according to claim 12 wherein R¹ is CN.
 15. A compound according to claim 12 wherein X¹ is CH₂.
 16. A compound according to claim 12 wherein X¹ is S.
 17. A compound according to claim 12 wherein b is
 1. 18. A compound according to claim 12 wherein b is
 2. 19. A compound according to claim 12 wherein a is
 1. 20. A compound according to claim 12 wherein X³, X⁴ and X⁵ are all N.
 21. A compound according to general formula 10, or a pharmaceutically acceptable salt thereof,

wherein: a is 0, 1 or 2; b is 1 or 2; X¹ is selected from CH₂, S, CF₂, CHF, CH(CH₃), C(CH₃)₂, CH(CN) and O; X² is selected from CR₂, O and S, provided that if a is 1 then X² is CH₂; X⁷ is selected from O, S, CH₂ and NH; R¹ is selected from H and CN; R⁹ is selected from H and alkyl; R¹⁰, R¹¹, R¹², R¹³ and R¹⁴ are independently selected from H, Br, Cl, F, CF₃, alkyl, acyl, OH, O-alkyl, NH₂, NH-alkyl, N(alkyl)₂, NO₂, NH-acyl, CO₂H, CO₂-alkyl, CONH₂, CONH-alkyl, CON(alkyl)₂ and CN is selected from H, Cl, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂.
 22. A compound according to claim 21 wherein R¹ is H.
 23. A compound according to claim 21 wherein R¹ is CN.
 24. A compound according to claim 21 wherein X¹ is CH₂.
 25. A compound according to claim 21 wherein X¹ is S.
 26. A compound according to claim 21 wherein b is
 1. 27. A compound according to claim 21 wherein b is
 2. 28. A compound according to claim 21 wherein a is
 1. 29. A compound according claim 21 wherein a is 2 and X² is CH₂.
 30. A compound according to general formula 11, or a pharmaceutically acceptable salt thereof,

wherein: a is 1 or 2; b is 1 or 2; X¹ is selected from CH₂, S, CF₂, CHF, CH(CH₃), C(CH₃)₂, CH(CN) and O; X⁷ is selected from O, S, CH₂ and NH; R¹ is selected from H and CN; R⁹ is selected from H and alkyl; R¹⁰, R¹¹, R¹², R¹³ and R¹⁴ are independently selected from H, Br, Cl, F, CF₃, alkyl, acyl, OH, O-alkyl, NH₂, NH-alkyl, N(alkyl)₂, NO₂, NH-acyl, CO₂H, CO₂-alkyl, CONH-alkyl, CON(alkyl)₂ and CN is selected from H, Cl, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂.
 31. A compound according to claim 30 wherein R¹ is H.
 32. A compound according to claim 30 wherein R¹ is CN.
 33. A compound according to claim 30 wherein X¹ is CH₂.
 34. A compound according to claim 30 wherein X¹ is S.
 35. A compound according to claim 30 wherein b is
 1. 36. A compound according to claim 30 wherein b is
 2. 37. A compound according to claim 30 wherein a is
 1. 38. A compound according to general formula 12, or a pharmaceutically acceptable salt thereof,

wherein: a is 0, 1 or 2; b is 1 or 2; X¹ is selected from CH₂, S, CF₂, CHF, CH(CH₃), C(CH₃)₂, CH(CN) and O; X² is selected from CH₂, O and S, provided that if a is l then X² is CH₂; R¹ is selected from H and CN; R¹⁵ and R¹⁶ are each independently selected from H, alkyl, alkenyl, polyfluoroalkyl, aralkyl, aryl and CH₂—L—R¹⁷; or R¹⁵ and R¹⁶ together are a group according to general formula 5, a group according to general formula 6 or a group according to general formula 7;

R¹⁷ is selected from H, alkyl and aryl; R¹⁸ is selected from H, alkyl, aryl, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂; R¹⁹ is selected from H, alkyl, aryl, F, Cl, Br, CF₃, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂; L is selected from a covalent bond, CH═CH, C≡C and —C₆H4—; d and e are selected from 0, 1, 2 and 3 such that d+e is 3,4 or 5; and f is selected from 1, 2 and 3; provided that when R¹⁵ and R¹⁶ are both H and b is 1 then X¹ is not S or CH₂.
 39. A compound according to claim 38 wherein R¹ is H.
 40. A compound according to claim 38 wherein R¹ is CN.
 41. A compound according to claim 38 wherein X¹ is CH₂.
 42. A compound according to claim 38 wherein X¹ is S.
 43. A compound according to claim 38 wherein b is
 1. 44. A compound according to claim 38 wherein b is
 2. 45. A compound according to claim 38 wherein a is
 1. 46. A compound according to claim 38 wherein a is 2 and X² is CH₂.
 47. A compound according to general formula 13, or a pharmaceutically acceptable salt thereof,

wherein: a is 1 or 2; b is 1 or 2; X¹ is selected from CH₂, S, CF₂, CHF, CH(CH₃), C(CH₃)₂, CH(CN) and O; R¹ is selected from H and CN; R¹⁵ and R¹⁶ are each independently selected from H, alkyl, alkenyl, polyfluoroalkyl, aralkyl, aryl and CH₂—L—R¹⁷; or R¹⁵ and R¹⁶ together are a group according to general formula 5, a group according to general Formula 6 or a group according to general formula 7;

R¹⁷ is selected from H, alkyl and aryl; R¹⁸ is selected from H, alkyl, aryl, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂; R¹⁹ is selected from H, alkyl, aryl, F, Cl, Br, CF₃, OH, O-alkyl, NH₂, NH-alkyl and N(alkyl)₂; L is selected from a covalent bond, CH═CH, C≡C and —C₆H₄—; d and e a reselected from 0, 1, 2 and 3 such that d+e is 3, 4 or 5; and f is selected from 1, 2 and
 3. 48. A compound according to claim 47 wherein R¹ is H.
 49. A compound according to claim 47 wherein R¹ is CN.
 50. A compound according to claim 47 wherein X1 is CH₂.
 51. A compound according to claim 47 wherein X¹ is S.
 52. A compound according to claim 47 wherein b is
 1. 53. A compound according to claim 47 wherein b is
 2. 54. A compound according to claim 47 wherein a is
 1. 55. A pharmaceutical composition comprising a compound according to claim
 1. 56. A use for a compound according to claim 1, which is as a component in the preparation of a pharmaceutical composition.
 57. A method of treatment of disease in a human or animal subject, comprising a step of administering to the subject a therapeutically active amount of a compound according to claim
 1. 58. A method of treatment according to claim 57 where the disease is caused by dysregulation of a post-proline cleaving proteases or their endogenous substrates.
 59. A method of treatment according to claim 57 where the disease is ameliorated by inhibition of a post-proline cleaving proteases.
 60. A method of treatment according to claim 57 where the disease is caused by dysregulation of a post-proline cleaving proteases or its endogenous substrates which is an intracellular protease.
 61. A composition according to claim 1 or with the proviso that when X¹=S; b=1; R¹=H; G²=H; G¹ is —CH₂—X²—(CH₂)_(a)—G³; a=1, X²=CH₂; G³=NR¹⁵R¹⁶; and one of R¹⁵, R¹⁶=H, the other of R¹⁵, R¹⁶ is not pyridyl, substituted pyridyl, pyrazinyl or substituted pyrazinyl.
 62. A composition according to claim 1, with the proviso that when b=1, R¹ is H and X¹ is S; G¹=H; G² is —CH₂—(CH₂)_(a)—G³; a=1; G³ is NR¹⁵R¹⁶ and one of R¹⁵ and R¹⁶ is H the other of R¹⁵, R¹⁶ is not pyridyl, substituted pyridyl, pyrazinyl or substituted pyrazinyl.
 63. A composition according to claim 1, with the proviso that when b=1, R¹ is CN and X¹ is CH₂; G¹=H; G² is —CH₂—(CH₂)_(a)—G³; a=1; G³ is NR¹⁵R¹⁶ and one of R¹⁵ and R¹⁶ is H, the other of R¹⁵, R¹⁶ is not pyridyl, substituted pyridyl, pyrazinyl or substituted pyrazinyl.
 64. A composition according to claim 1, with the proviso that when G²=H; G¹=—CH₂—X²—(CH₂)_(a) 13 G³; X² is CH₂; a=1; G³=NR¹⁵R¹⁶ and R¹⁵=R¹⁶=H; b is not 2 when X¹ is O or CH₂, and b is not 1 when X¹ is CH₂.
 65. A method of treatment according to claim 57 in which the disease is caused by dysregulation of a non-membrane associated post-proline cleaving proteases such as QPP, DPP-8 and DPP-9 enzymes or their endogenous substrates.
 66. A method of treatment according to claim 57 in which the disease is ameliorated by inhibition of a non-membrane associated post-proline cleaving proteases such as QPP, DPP-8 and DPP-9 enzymes or their endogenous substrates.
 67. A method claim 57 in which the compound is a selective inhibitor of non-membrane associated post-proline cleaving proteases. 